Aryldiazepine derivatives as RSV inhibitors

ABSTRACT

The present invention discloses compounds of Formula (I), and pharmaceutically acceptable salts, esters, or prodrugs thereof: 
                         
which inhibit Respiratory Syncytial Virus (RSV). The present invention further relates to pharmaceutical compositions comprising the aforementioned compounds for administration to a subject suffering from RSV infection. The invention also relates to methods of treating an RSV infection in a subject by administering a pharmaceutical composition comprising the compounds of the present invention.

RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.62/516,274, filed on Jun. 7, 2017. The entire teachings of the aboveapplication are incorporated herein by reference.

TECHNICAL FIELD

The present invention relates generally to compounds and pharmaceuticalcompositions useful as Respiratory Syncytial Virus (RSV) inhibitors.Specifically, the present invention relates to aryldiazepine derivativesthat can inhibit RSV activities and for treating RSV infection.

BACKGROUND OF THE INVENTION

Human respiratory syncytial virus (HRSV) is a negative-sense, singlestranded, RNA paramyxovirus (K M. Empey, et al., Rev. Anti-InfectiveAgents, 2010, 50 (1 May), 1258-1267). RSV is the leading cause of acutelower respiratory tract infections (ALRI) and affects patients of allages. The symptoms in adults are usually not severe and are typicallyanalogous to a mild cold. However, in infants and toddlers the virus cancause lower respiratory tract infections including bronchiolitis orpneumonia with many of them requiring hospitalization. Nearly allchildren have been infected by age 3. There are known high-risk groupsthat infection with RSV is more likely to progress into the ALRI.Premature infants and/or infants suffering from lung or cardiac diseaseare at the highest risk to develop ALRI. Additional high-risk groupsinclude the elderly, adults with chronic heart and/or lung disease, stemcell transplant patients and the immunosuppressed.

Currently, there is no vaccine available to prevent HRSV infection.Palivizumab is a monoclonal antibody that is used prophylactically toprevent HRSV infection in high risk infants, e.g. premature infants, andinfants with cardiac and/or lung disease. The high cost of palivizumabtreatment limits its use for general purposes. Ribavirin has also beenused to treat HRSV infections but its effectiveness is limited. There isa major medical need for new and effective HRSV treatments that can beused generally by all population types and ages.

There have been several RSV fusion inhibitors that have been disclosedin the following publications: WO2010/103306, WO2012/068622,WO2013/096681, WO2014/060411, WO2013/186995, WO2013/186334, WO2013/186332, WO 2012 080451, WO 2012/080450, WO2012/080449, WO2012/080447, WO 2012/080446, and J. Med. Chem. 2015, 58, 1630-1643.Examples of other N-protein inhibitors for treatment of HRSV have beendisclosed in the following publications: WO 2004/026843, J. Med. Chem.2006, 49, 2311-2319, and J. Med. Chem. 2007, 50, 1685-1692. Examples ofL-protein inhibitors for HRSV have been disclosed in the followingpublications: WO 2011/005842, WO 2005/042530, Antiviral Res. 2005, 65,125-131, and Bioorg. Med. Chem. Lett. 2013, 23, 6789-6793. Examples ofnucleosides/polymerase inhibitors have been disclosed in the followingpublications: WO 2013/242525 and J. Med. Chem. 2015, 58, 1862-1878.

There is a need for the development of effective treatments for HRSV.The present invention has identified these novel compounds and theirinhibitory activity against HRSV. The invention includes methods toprepare the compounds as well as methods of using these compounds totreat disease.

SUMMARY OF THE INVENTION

The present invention provides compounds represented by Formula (I), andpharmaceutically acceptable salts, esters and prodrugs thereof:

wherein:A is aryl or heteroaryl, each of which, when possible, is optionallysubstituted with one or more substituents which are not R₃;B is optionally substituted heteroaryl or optionally substituted aryl;L is —NH—, —NHC(O)—, or —NHC(O)NH—;X is —CH₂—, or —C(O)—;R₁ is optionally substituted heteroaryl or optionally substituted aryl;R₂ is absent or selected from the group consisting of:

1) Optionally substituted —C₁-C₈ alkyl;

2) Optionally substituted —C₂-C₈ alkenyl;

3) Optionally substituted —C₂-C₈ alkynyl;

4) Optionally substituted —C₁-C₈ alkoxy;

5) Optionally substituted aryloxy;

6) Optionally substituted —C₃-C₁₂ cycloalkyl;

7) Optionally substituted —C₃-C₁₂ cycloalkenyl;

8) Optionally substituted 3- to 12-membered heterocycloalkyl;

9) Optionally substituted aryl;

10) Optionally substituted arylalkyl;

11) Optionally substituted heteroaryl;

12) Optionally substituted heteroarylalkyl;

13) —NR₁₃R₁₄;

14) —CO—NR₁₃R₁₄; and

15) —SO₂—NR₁₃R₁₄;

-   -   preferably, R₂ is optionally substituted —C₃-C₁₂ cycloalkyl;        optionally substituted —C₃-C₁₂ cycloalkenyl; optionally        substituted 3- to 12-membered heterocycloalkyl; optionally        substituted aryl; or optionally substituted heteroaryl;    -   each R₃ is the same or different and independently selected from        halogen, hydroxyl, protected hydroxyl, cyano, amino, protected        amino, nitro, optionally substituted —C₁-C₈ alkyl, optionally        substituted —C₁-C₈ alkoxy, optionally substituted —NHC₁-C₈        alkyl, optionally substituted —S—(—C₁-C₈ alkyl), optionally        substituted —SO₂—(—C₁-C₈ alkyl), -optionally substituted        —SO₂—NH—(—C₁-C₈ alkyl), optionally substituted —NH—SO₂—(—C₁-C₈        alkyl), —CO₂R₁₂, —NR₁₃R₁₄, and —CO—NR₁₃R₁₄;        R₁₂ is independently selected from the group consisting of:

1) Optionally substituted —C₁-C₈ alkyl;

2) Optionally substituted —C₂-C₈ alkenyl;

3) Optionally substituted —C₂-C₈ alkynyl;

4) Optionally substituted —C₃-C₈ cycloalkyl;

5) Optionally substituted —C₃-C₈ cycloalkenyl;

6) Optionally substituted 3- to 8-membered heterocycloalkyl;

7) Optionally substituted aryl; and

8) Optionally substituted heteroaryl;

-   -   R₁₃ and R₁₄ are each independently selected from hydrogen,        optionally substituted —C₁-C₈-alkyl, optionally substituted        —C₂-C₈-alkenyl, optionally substituted —C₂-C₈-alkynyl;        optionally substituted —C₃-C₈-cycloalkyl, optionally substituted        —C₃-C₈ cycloalkenyl; optionally substituted 3- to 12-membered        heterocycloalkyl, optionally substituted aryl; optionally        substituted heteroaryl; optionally substituted —C₁-C₈-alkoxy,        —C(O)R₁₂, —S(O)₂R₁₂, and —S(O)₂NHR₁₂; alternatively, R₁₃ and R₁₄        are taken together with the nitrogen atom to which they are        attached to form an optionally substituted heterocyclic ring;

R₄ is selected from hydrogen and optionally substituted —C₁-C₈ alkyl,preferably R₄ is hydrogen or CH₃; and

n is 0 to k, wherein k is the total number of CH and NH groups in theheteroaryl group A when A is unsubstituted; preferably, n is 0, 1 or 2;more preferably n is 0. Most preferably, A is unsubstituted.

Each preferred group stated above can be taken in combination with one,any or all other preferred groups.

DETAILED DESCRIPTION OF THE INVENTION

In one embodiment of the present invention is a compound represented byFormula (I) as described above, or a pharmaceutically acceptable salt,ester or prodrug thereof.

The carbon atom at position 3 of the aryldiazepine ring system of thecompounds of the invention is chiral. Thus, compounds of the inventioncan have the stereochemistry depicted in Formula (Ia) or (Ib):

wherein R₁, R₂, R₃, R₄, A, B, L, X, and n are as previously defined. Acomposition of the invention can comprise a compound of the invention asa racemic mixture of Formula Ia and Formula Ib, a pure enantiomer ofeither Formula Ia or Formula Ib, or an excess of one enantiomer over theother. For example, the composition can comprise the compound in anenantiomeric excess of at least 5, 10, 20, 30, 40, 50, 60, 70, 80 or90%. In one embodiment, the enantiomeric excess is at least 95%. Incompounds of the invention having two or more chiral atoms, suchcompounds can be present in a composition as a pure stereoisomer or amixture of stereoisomers, such as a racemic mixture or a mixture ofdiasteromers. In one embodiment, a composition of the inventioncomprises a racemic mixture, a single stereoisomer or enantiomers withan enantiomeric excess of at least 5, 10, 20, 30, 40, 50, 60, 70, 80, 90or 95%.

In a preferred embodiment, a compound of the invention is represented byFormula (Ib). Compositions of the invention preferably comprise asubstantially pure compound of Formula (Ib), or a mixture of a compoundof Formula (Ib) and the corresponding compound of Formula (Ia), with anenantiomeric excess of the compound of Formula (Ib) as discussed above.

In certain embodiments, the present invention relates to compounds ofFormula (I), and pharmaceutically acceptable salts thereof, wherein A isoptionally substituted aryl. Preferably A is optionally substitutedphenyl, more preferably A is unsubstituted phenyl.

In certain embodiments, the present invention relates to compounds ofFormula (I), and pharmaceutically acceptable salts thereof, wherein A isoptionally substituted heteroaryl.

Preferably A is optionally substituted heteroaryl. More preferably A isoptionally substituted monocyclic 5-membered heteroaryl. In oneembodiment, A is a five-membered heteroaryl group containing sulfur.

In another embodiment, the present invention relates to compounds ofFormula (I), and pharmaceutically acceptable salts thereof, wherein A isderived from one of the following by removal of the hydrogen atoms fromtwo adjacent carbon atoms:

each of which, when possible, is optionally substituted with one or moresubstituents which are not R₃.

In certain embodiments, the present invention relates to compounds ofFormula (I), and pharmaceutically acceptable salts thereof, wherein eachR₃ is independently halo, optionally substituted —C₁-C₄-alkyl, or—C₁-C₄-alkoxy. In certain embodiments, the present invention relates tocompounds of Formula (I), and pharmaceutically acceptable salts thereof,wherein each R₃ is independently —F, —CH₃, —CF₃, —OCF₃, —CN, —SO₂Me, or—CH₂N(CH₃)₂. In certain embodiments, the present invention relates tocompounds of Formula (I), and pharmaceutically acceptable salts thereof,wherein n is 0 to 3, 0 to 2, 1 or 0. More preferably, n is 0.

In certain embodiments, the present invention relates to compounds ofFormula (I), and pharmaceutically acceptable salts thereof, wherein R₁is optionally substituted aryl. In certain embodiments, the presentinvention relates to compounds of Formula (I), and pharmaceuticallyacceptable salts thereof, wherein R₁ is optionally substitutedheteroaryl.

In certain embodiments, the present invention relates to compounds ofFormula (I), and pharmaceutically acceptable salts thereof, wherein R₁is phenyl optionally substituted with one to three substituents selectedfrom the group consisting of halo, —CF₃, —OCF₃, —CH₃, —SO₂Me, and cyano.Preferably, R₁ is unsubstituted phenyl.

In one embodiment, the present invention relates to compounds of Formula(I), and pharmaceutically acceptable salts thereof, wherein R₄ isoptionally substituted methyl. In another embodiment, the presentinvention relates to compounds of Formula (I), and pharmaceuticallyacceptable salts thereof, wherein R₄ is hydrogen.

In one embodiment, the present invention relates to compounds of Formula(I), and pharmaceutically acceptable salts thereof, wherein X is —CH₂—.In another embodiment, the present invention relates to compounds ofFormula (I), and pharmaceutically acceptable salts thereof, wherein X is—C(O)—.

In one embodiment, the present invention relates to compounds of Formula(I), and pharmaceutically acceptable salts thereof, wherein L is —NH—.In another embodiment, the present invention relates to compounds ofFormula (I), and pharmaceutically acceptable salts thereof, wherein L is—NHC(O)—. In another embodiment, the present invention relates tocompounds of Formula (I), and pharmaceutically acceptable salts thereof,wherein L is —NHC(O)NH—.

In certain embodiments, the present invention relates to compounds ofFormula (I), and pharmaceutically acceptable salts thereof, wherein B isoptionally substituted aryl, optionally substituted heteroaryl.Preferably B is optionally substituted monocyclic 5-membered heteroaryl,a monocyclic 6-membered heteroaryl or an 8-10-membered fused heteroaryl.In one embodiment, B is a five-membered nitrogen containing heteroarylgroup.

In another embodiment, the present invention relates to compounds ofFormula (I), and pharmaceutically acceptables salt thereof, wherein B isderived from one of the following by removal of two hydrogen atoms:

wherein each of the above is optionally substituted when possible.

In certain embodiments, B is selected from, but not limited to, thegroups set forth below, where one of the indicated valences is the pointof attachment of the heteroaryl group to R₂ and the other is the pointof attachment to L. Each of these groups is optionally additionallysubstituted when possible. The atom of B which connects to L is a carbonatom. The atom of B that connects to R₂ is a carbon atom or, whenpossible, a nitrogen atom:

In another particular embodiment, the present invention relates tocompounds of Formula (I), and pharmaceutically acceptable salts thereof,wherein B is derived from a fused bicyclic group selected from one ofthe following by removal of two hydrogen atoms.

In certain embodiments, the present invention relates to compounds ofFormula (I), and pharmaceutically acceptable salts thereof, where B isselected from the groups set forth below,

wherein the point of attachment to L is indicated and R₂ is attached toany other available ring position. In one embodiment, R₂ is attached toan atom of the benzo ring. When B is naphthyl, R₂ and L are preferablyattached to carbon atoms from different rings. Each of the above showngroups is optionally substituted, and preferably the optionalsubstituents are independently selected from halo, —CH₃, —CF₃, —OCF₃,—CN, —NH₂, —OH, —CH₂N(CH₃)₂, —C(O)CH₃, —NH—(C₁-C₆)alkyl,—SO₂—(C₁-C₆)alkyl, —SO₂—NH—(C₁-C₆)alkyl, —NH—SO₂—(C₁-C₆)alkyl, and—C₁-C₈-alkoxy. Preferably, in addition to R₂, there are 0, 1, 2 or 3substituents, more preferably 0, 1 or 2 substituents, and mostpreferably 0 or 1 substituent.

In certain embodiments of the compounds of the invention, R₂ is absent.

In certain embodiments, R₂ is an optionally substituted aryl,heteroaryl, 3- to 12-membered heterocycloalkyl, —C₃-C₁₂-cycloalkyl,—C₃-C₁₂ cycloalkenyl, aryl-O—, heteroaryl-O, aryl-C₁-C₄-alkyl orheteroaryl-C₁-C₄-alkyl. In certain embodiments, R₂ is phenyl, naphthyl,5-membered heteroaryl or 6-membered heteroaryl, each of which isoptionally substituted. In certain embodiments, R₂ is a 5- or 6-memberedheteroaryl fused with a 6-membered aryl, heteroaryl, carbocyclic orheterocyclic ring, such as a benzo-fused-5- or 6-membered heteroaryl ora pyrido-fused 5- or 6-membered heteroaryl.

In certain embodiments of the compounds of the invention, R₂ is a groupderived from one of the following by removal of one hydrogen atom from aring atom:

wherein each of the above shown is optionally substituted when possible.

In certain embodiments, R₂ is selected from the groups shown below, eachof which is optionally substituted,

In certain embodiments, R₂ is optionally substituted with one or moresubstituents independently selected from halo, —CH₃, —CF₃, —OCF₃, —CN,—NH₂, —OH, —CH₂N(CH₃)₂, —C(O)CH₃, optionally substituted—NH—(C₁-C₆)alkyl, optionally substituted —NH—(C₁-C₆)alkyl-(C₁-C₆)alkoxy,optionally substituted —SO₂—(C₁-C₆)alkyl, optionally substituted—SO₂—NH—(C₁-C₆)alkyl, optionally substituted —NH—SO₂—(C₁-C₆)alkyl,optionally substituted 3- to 12-membered heterocycloalkyl, optionallysubstituted aryl, optionally substituted heteroaryl, optionallysubstituted —C₁-C₈-alkyl, optionally substituted —C₁-C₈-alkenyl,optionally substituted —C₃-C₈-cycloalkyl, optionally substituted—C₃-C₈-cycloalkenyl, and optionally substituted —C₁-C₈-alkoxy. Incertain embodiments, there are 0 to 4, 0 to 3, 0 to 2, 1 or 0substituents. Preferably, there are 0 to 2 substituents and morepreferably, 0 or 1 substituent. More preferably optionally substitutedgroups can be more than one. In this embodiment, R₂ is optionallysubstituted with one or more aryl, heteroaryl, 3- to 12-memberedheterocycloalkyl, C₃-C₁₂-cycloalkyl, C₃-C₁₂ cycloalkenyl, aryl-O—,heteroaryl-O, aryl-C₁-C₄-alkyl or heteroaryl-C₁-C₄-alkyl, as describedabove. In this embodiment, R₂ is preferably optionally substituted aryl,heteroaryl, 3- to 12-membered heterocycloalkyl, C₃-C₁₂-cycloalkyl, orC₃-C₁₂ cycloalkenyl.

In certain embodiments of the compounds of the invention, R₂ is selectedfrom the groups below, each of which is optionally substituted:

In certain embodiments of the compounds of the invention, R₂ is selectedfrom the groups below, each of which is optionally substituted:

In certain embodiments of the compounds of the invention, R₂ isoptionally substituted with one or more substituents independentlyselected from the groups below, each of which can be optionallysubstituted further:

In certain embodiments of the compounds of the invention, A is aoptionally substituted monocyclic 5-membered heteroaryl; R₁ isoptionally substituted aryl; R₂ is optionally substituted aryl oroptionally substituted heteroaryl; R₄ is methyl or hydrogen; and L is—NH—. Preferably A is optionally substituted thiophene. Preferably R₁ isoptionally substituted phenyl. Preferably B is optionally substitutedtriazole, optionally substituted oxadiazolyl, optionally substitutedoxazolyl, or optionally substituted thiadiazolyl.

In certain embodiments of the compounds of the invention, A isoptionally substituted aryl; R₁ is optionally substituted aryl; R₂ isoptionally substituted aryl or optionally substituted heteroaryl; R₄ ismethyl or hydrogen; and L is —NH—. Preferably A is optionallysubstituted phenyl. Preferably R₁ is optionally substituted phenyl.Preferably B is optionally substituted triazolyl, optionally substitutedoxadiazolyl, optionally substituted oxazolyl, or optionally substitutedthiadiazolyl.

In another embodiment, the invention provides a compound represented byone of Formulas (II-1)˜(II-3), (IIa-1)˜(IIa-3), and (IIb-1)˜(IIb-3) or apharmaceutically acceptable salt, ester or prodrug thereof:

wherein R₁, R₂, R₃, R₄, A, B, X, and n are as previously defined.

In another embodiment, the invention provides a compound represented byone of Formulas (III-1) and (III-2), or a pharmaceutically acceptablesalt, ester or prodrug thereof:

wherein R₁, R₂, R₃, R₄, A, B, L, and n are as previously defined.

In another embodiment, the invention provides a compound represented byone of Formulas (IV-1) and (IV-2), or a pharmaceutically acceptablesalt, ester or prodrug thereof:

wherein R₂, R₃, R₄, A, B, L, and n are as previously defined.

In another embodiment, the invention provides a compound represented byone of Formulas (V-1) and (V-2), or a pharmaceutically acceptable salt,ester or prodrug thereof:

wherein R₂, R₃, B, L, and n are as previously defined.

In another embodiment, the invention provides a compound represented byone of Formulas (VI-1)˜(VI-8), (VIa-1)˜(VIa-8), and (VIb-1)˜(VIb-8), ora pharmaceutically acceptable salt, ester or prodrug thereof:

wherein R₂, R₃, R₄, A, X, and n are as previously defined. Preferably R₄is hydrogen, A is phenyl, and n is 0.

In another embodiment of the invention is a compound represented by oneof Formulas (VII-1)˜(VII-8), (VIIa-1)˜(VIIa-8), and (VIIb-1)˜(VIIb-8) ora pharmaceutically acceptable salt, ester or prodrug thereof:

wherein R₂, R₃, R₄, A, X, m and n are as previously defined. PreferablyR₄ is hydrogen, A is phenyl, and n is 0.

In another embodiment, the invention provides compounds represented byFormulas (VIII-1)˜(VIII-8), (VIIIa-1)˜(VIIIa-8), and (VIIIb-1)˜(VIIIb-8)and pharmaceutically acceptable salts, esters and prodrugs thereof:

wherein R₂, R₃, R₄, A, X, m and n are as previously defined. PreferablyR₄ is hydrogen, A is phenyl, and n is 0.

In particular embodiments, the invention provides compounds of Formulas(VI-1)˜(VI-8), (VIa-1)˜(VIa-8), (VIb-1)˜(VIb-8), (VII-8),(VIIa-1)˜(VIIa-8), (VIIb-1)˜(VIIb-8), (VIII-8), (VIIIa-1)˜(VIIIa-8), and(VIIIb-1)˜(VIIIb-8) and pharmaceutically acceptable salts, esters andprodrugs thereof, where R₂ is selected from the groups set forth inTable 1 (Entry 1 to Entry 186 in Table 1), each of which is optionallyfurther substituted.

TABLE 1 Entry R₂ 1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

27

28

29

30

31

32

33

34

35

36

37

38

39

40

41

42

43

44

45

46

47

48

49

50

51

52

53

54

55

56

57

58

59

60

61

62

63

64

65

66

67

68

69

70

71

72

73

74

75

76

77

78

79

80

81

82

83

84

85

86

87

88

89

90

91

92

93

94

95

96

97

98

99

100

101

102

103

104

105

106

107

108

109

110

111

112

113

114

115

116

117

118

119

120

121

122

123

124

125

126

127

128

129

130

131

132

133

134

135

136

137

138

139

140

141

142

143

144

145

146

147

148

149

150

151

152

153

154

155

156

157

158

159

160

161

162

163

164

165

166

167

168

169

170

171

172

173

174

175

176

177

178

179

180

181

182

183

184

185

186

It will be appreciated that the description of the present inventionherein should be construed in congruity with the laws and principles ofchemical bonding. In some instances it may be necessary to remove ahydrogen atom in order to accommodate a substituent at any givenlocation.

It is intended that the definition of any substituent or variable (e.g.,R₁, R₂, etc.) at a particular location in a molecule be independent ofits definitions elsewhere in that molecule.

It will be yet appreciated that the compounds of the present inventionmay contain one or more asymmetric carbon atoms and may exist inracemic, diastereoisomeric, and optically active forms. It will still beappreciated that certain compounds of the present invention may exist indifferent tautomeric forms. All tautomers are contemplated to be withinthe scope of the present invention.

In certain embodiments, the present invention provides a method for theprevention or treatment of RSV activities and for treating RSV infectionis subjects. The method comprises administering a therapeuticallyeffective amount of a compound of formula (I).

The present invention also provides the use of a compound of formula (I)for the preparation of a medicament for the prevention or treatment ofRSV.

Thus, in one embodiment, a compound of formula (I), or pharmaceuticallyacceptable salt thereof, is combined with a steroid anti-inflammatorycompound, for example budesonide or fluticasone. In a preferredembodiment, the steroid is administered in low doses to minimizeimmuno-suppressant effects. In another embodiment a compound of formula(I), or a pharmaceutically acceptable salt thereof, is combined with anon-steroid anti-inflammatory compound, for example leukotrieneantagonists such as Singulair (Merck) or Accolate (Astra Zeneca),phosphodiesterase 4 inhibitors such as roflumilast (Altana), TNF alphainhibitors such as Enbrel (Amgen), Remicade (Centocor), Humira (Abbott)or CDP870 (Celltech) or NSAIDS. In a further embodiment, a compound offormula (I) is combined with interleukin 8 or interleukin 9 inhibitors.The present invention thus also relates to a product containing acompound of formula (I), or a pharmaceutically acceptable salt thereof,and an anti-inflammatory compound for simultaneous, separate orsequential use in the treatment of RSV.

The present invention also relates to a combination of a compound offormula (I), or a pharmaceutically acceptable salt thereof, with ananti-influenza compound and the use of such a combination in thetreatment of concomitant RSV and influenza infections. The presentinvention thus also relates to a product containing a compound offormula (I), or a pharmaceutically acceptable salt thereof, and ananti-influenza compound for simultaneous, separate or sequential use inthe treatment of concomitant RSV and influenza infections. The compoundsof the invention may be administered in a variety of dosage forms. Thus,they can be administered orally, for example as tablets, troches,lozenges, aqueous or oily suspensions, dispersible powders or granules.The compounds of the invention may also be administered parenterally,whether subcutaneously, intravenously, intramuscularly, intrasternally,transdermally or by infusion techniques. The compounds may also beadministered as suppositories.

In an embodiment, the compounds of the invention are administered byintranasal or intrabronchial administration. The present invention alsoprovides an inhaler or nebuliser containing a medicament which comprises(a) a benzodiazepine derivative of the formula (I), as defined above, ora pharmaceutically acceptable salt thereof, and (b) a pharmaceuticallyacceptable carrier or diluent.

The present invention also provides a pharmaceutical compositioncontaining such a benzodiazepine derivative, or a pharmaceuticallyacceptable salt thereof, and a pharmaceutically acceptable carrier ordiluent.

The compounds of the invention are typically formulated foradministration with a pharmaceutically acceptable carrier or diluent.For example, solid oral forms may contain, together with the activecompound, diluents, e.g. lactose, dextrose, saccharose, cellulose, cornstarch or potato starch; lubricants, e.g. silica, talc, stearic acid,magnesium or calcium stearate, and/or polyethylene glycols; bindingagents; e.g. starches, arabic gums, gelatin, methylcellulose,carboxymethylcellulose or polyvinyl pyrrolidone; disaggregating agents,e.g. starch, alginic acid, alginates or sodium starch glycolate;effervescing mixtures; dyestuffs; sweeteners; wetting agents, such aslecithin, polysorbates, laurylsulphates; and, in general, non toxic andpharmacologically inactive substances used in pharmaceuticalformulations. Such pharmaceutical preparations may be manufactured inknown manner, for example, by means of mixing, granulating, tableting,sugar coating, or film coating processes.

Liquid dispersions for oral administration may be syrups, emulsions andsuspensions. The syrups may contain as carriers, for example, saccharoseor saccharose with glycerine and/or mannitol and/or sorbitol.

Suspensions and emulsions may contain as carrier, for example a naturalgum, agar, sodium alginate, pectin, methylcellulose,carboxymethylcellulose, or polyvinyl alcohol. The suspension orsolutions for intramuscular injections may contain, together with theactive compound, a pharmaceutically acceptable carrier, e.g. sterilewater, olive oil, ethyl oleate, glycols, e.g. propylene glycol, and ifdesired, a suitable amount of lidocaine hydrochloride.

Solutions for injection or infusion may contain as carrier, for example,sterile water or preferably they may be in the form of sterile, aqueous,isotonic saline solutions.

The present invention also relates to the novel compounds, as definedabove, for use in a method of treating the human or animal body. Thepresent invention also relates to a pharmaceutical compositioncomprising a novel compound as defined above and a pharmaceuticallyacceptable diluant or carrier. Preferably, the pharmaceuticalcomposition comprises a pharmaceutically acceptable salt of a novelcompound as defined above. A pharmaceutically acceptable salt is asdefined above. The novel compounds of the invention are typicallyadministered in the manner defined above and the compounds are typicallyformulated for administration in the manner defined above.

Preferably, the pharmaceutical compositions comprise optically activeisomers of the novel compounds of the invention. Thus, for example,preferred novel compounds of the invention containing only one chiralcentre include an R enantiomer in substantially pure form, an Senantiomer in substantially pure form and enantiomeric mixtures whichcontain an excess of the R enantiomer or an excess of the S enantiomer.It is particularly preferred that a pharmaceutical composition containsa compound of the invention which is a substantially pure opticalisomer. For the avoidance of doubt, the novel compounds of the inventioncan, if desired, be used in the form of solvates.

Yet a further aspect of the present invention is a process of making anyof the compounds delineated herein employing any of the synthetic meansdelineated herein.

Definitions

Listed below are definitions of various terms used to describe thisinvention. These definitions apply to the terms as they are usedthroughout this specification and claims, unless otherwise limited inspecific instances, either individually or as part of a larger group.

The term “alkyl”, as used herein, refers to a saturated, monovalentstraight- or branched-chain hydrocarbon radicals. Preferred alkylradicals include C₁-C₆ alkyl and C₁-C₈ alkyl radicals. Examples of C₁-C₆alkyl radicals include, but are not limited to, methyl, ethyl, propyl,isopropyl, n-butyl, tert-butyl, neopentyl, n-hexyl radicals; andexamples of C₁-C₈ alkyl groups include, but are not limited to, methyl,ethyl, propyl, isopropyl, n-butyl, tert-butyl, neopentyl, n-hexyl,heptyl, and octyl radicals.

The term “alkenyl” as used herein, refers to straight- or branched-chainhydrocarbon radicals having at least one carbon-carbon double bond bythe removal of a single hydrogen atom. Preferred alkenyl groups includeC₂-C₆ alkenyl and C₂-C₈ alkenyl groups. Alkenyl groups include, but arenot limited to, for example, ethenyl, propenyl, butenyl,1-methyl-2-buten-1-yl, heptenyl, octenyl and the like.

The term “alkynyl” as used herein, refers to straight- or branched-chainhydrocarbon radicals having at least one carbon-carbon triple bond bythe removal of a single hydrogen atom. Preferred alkynyl radicalsinclude C₂-C₆ alkynyl and C₂-C₈ alkynyl radicals. Representative alkynylradicals include, but are not limited to, for example, ethynyl,1-propynyl, 1-butynyl, heptynyl, octynyl and the like.

It is understood that any alkyl, alkenyl, alkynyl and cycloalkyl moietydescribed herein can also be an aliphatic group, an alicyclic group or aheterocyclic group. An “aliphatic” group is a non-aromatic moiety thatmay contain any combination of carbon atoms, hydrogen atoms, halogenatoms, oxygen, nitrogen or other atoms, and optionally contain one ormore units of unsaturation, e.g., double and/or triple bonds. Analiphatic group may be straight chained, branched or cyclic andpreferably contains between about 1 and about 24 carbon atoms, moretypically between about 1 and about 12 carbon atoms. In addition toaliphatic hydrocarbon groups, aliphatic groups include, for example,polyalkoxyalkyls, such as polyalkylene glycols, polyamines, andpolyimines, for example. Such aliphatic groups may be furthersubstituted.

The term “alicyclic,” as used herein, denotes a monovalent group derivedfrom a monocyclic or bicyclic saturated carbocyclic ring compound by theremoval of a single hydrogen atom. Examples include, but not limited to,cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, bicyclo [2.2.1]heptyl,and bicyclo [2.2.2] octyl. Such alicyclic groups may be furthersubstituted.

The term “alkynylene” refers to an alkynyl group from which anadditional hydrogen atom has been removed to form a diradical group.Alkynylene groups include, but are not limited to, for example,ethynylene, propynylene, butynylene, 1-methyl-2-butyn-1-ylene,heptynylene, octynylene, and the like.

The term “carbocycle” refers to a saturated (e.g., “cycloalkyl”),partially saturated (e.g., “cycloalkenyl” or “cycloalkynyl”) orcompletely unsaturated (e.g., “aryl”) ring system containing zeroheteroatom ring atom. “Ring atoms” or “ring members” are the atoms boundtogether to form the ring or rings. Where a carbocycle group is adivalent moiety linking two other elements in a depicted chemicalstructure, the carbocycle group can be attached to the two otherelements through any two substitutable ring atoms. A C₄-C₆ carbocyclehas 4-6 ring atoms.

The term “cycloalkyl”, as used herein, refers to a monocyclic orpolycyclic saturated carbocyclic ring compound, and the carbon atoms maybe optionally oxo-substituted. A polycyclic cycloalkenyl can comprisefused rings. Preferred cycloalkyl groups include C₃-C₈ cycloalkyl andC₃-C₁₂ cycloalkyl groups. Examples of C₃-C₈-cycloalkyl include, but arenot limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cyclopentyl and cyclooctyl; and examples of C₃-C₁₂-cycloalkyl include,but not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,bicyclo [2.2.1] heptyl, and bicyclo [2.2.2] octyl.

The term “cycloalkenyl”, as used herein, refers to monocyclic orpolycyclic carbocyclic ring compound having at least one carbon-carbondouble bond and the carbon atoms may be optionally oxo-substituted. Apolycyclic cycloalkenyl can comprise fused rings, covalently attachedrings or a combination thereof. Preferred cycloalkenyl groups includeC₃-C₈ cycloalkenyl and C₃-C₁₂ cycloalkenyl groups. Examples ofC₃-C₈-cycloalkenyl include, but are not limited to, cyclopropenyl,cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl,and the like; and examples of C₃-C₁₂-cycloalkenyl include, but notlimited to, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl,cycloheptenyl, cyclooctenyl, and the like.

The terms “heterocyclic” or “heterocycloalkyl” can be usedinterchangeably and referred to a non-aromatic ring or a bi- ortri-cyclic group fused, bridged or spiro system, where (i) each ringsystem contains at least one heteroatom independently selected fromoxygen, sulfur and nitrogen, (ii) each ring system can be saturated orunsaturated (iii) the nitrogen and sulfur heteroatoms may optionally beoxidized, (iv) the nitrogen heteroatom may optionally be quaternized,(v) any of the above rings may be fused to an aromatic ring, and (vi)the remaining ring atoms are carbon atoms which may be optionallyoxo-substituted or optionally substituted with exocyclic olefinic,iminic or oximic double bond. Representative heterocycloalkyl groupsinclude, but are not limited to, 1,3-dioxolane, pyrrolidinyl,pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl,piperazinyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl,isothiazolidinyl, quinoxalinyl, pyridazinonyl,2-azabicyclo[2.2.1]-heptyl, 8-azabicyclo[3.2.1]octyl,5-azaspiro[2.5]octyl, 1-oxa-7-azaspiro[4.4]nonanyl, 7-oxooxepan-4-yl,and tetrahydrofuryl. Such heterocyclic groups may be furthersubstituted. Heteroaryl or heterocyclic groups can be C-attached orN-attached (where possible).

The term “aryl,” as used herein, refers to a mono- or polycycliccarbocyclic ring system comprising at least one aromatic ring,including, but not limited to, phenyl, naphthyl, tetrahydronaphthyl,indanyl, indenyl and the like. A polycyclic aryl is a polycyclic ringsystem that comprises at least one aromatic ring. Polycyclic aryls cancomprise fused rings, covalently attached rings or a combinationthereof.

The term “heteroaryl,” as used herein, refers to a mono- or polycyclicaromatic radical having one or more ring atom selected from S, O and N;and the remaining ring atoms are carbon, wherein any N or S containedwithin the ring may be optionally oxidized. Preferred heteroaryl groupsare monocyclic or bicyclic. Heteroaryl groups include, but are notlimited to, pyridinyl, pyrazinyl, pyrimidinyl, pyrrolyl, pyrazolyl,imidazolyl, thiazolyl, oxazolyl, isooxazolyl, thiadiazolyl, oxadiazolyl,thiophenyl, furanyl, quinolinyl, isoquinolinyl, benzimidazolyl,benzooxazolyl, quinoxalinyl, and the like. A polycyclic heteroaryl cancomprise fused rings, covalently attached rings or a combinationthereof.

In accordance with the invention, aromatic groups can be substituted orunsubstituted.

The term “arylalkyl,” as used herein, refers to functional group whereinan alkylene chain is attached to an aryl group. Examples include, butare not limited to, benzyl, phenethyl and the like. The term“substituted arylalkyl” means an arylalkyl functional group in which thearyl group is substituted. Similarly, the term “heteroarylalkyl” means afunctional group wherein an alkylene chain is attached to a heteroarylgroup. Examples include, but are not limited to, pyridinylmethyl,pyrimidinylethyl and the like. The term “substituted heteroarylalkyl”means a heteroarylalkyl functional group in which the heteroaryl groupis substituted.

The term “alkoxy” employed alone or in combination with other termsmeans, unless otherwise stated, an alkyl group having the designatednumber of carbon atoms connected to the rest of the molecule via anoxygen atom, such as, for example, methoxy, ethoxy, 1-propoxy, 2-propoxy(isopropoxy) and the higher homologs and isomers. Preferred alkoxy are(C₁-C₃) alkoxy.

The term “halo” or halogen” alone or as part of another substituent, asused herein, refers to a fluorine, chlorine, bromine, or iodine atom.

The term “hydrogen” includes hydrogen and deuterium. In addition, therecitation of an atom includes other isotopes of that atom so long asthe resulting compound is pharmaceutically acceptable.

The term “substituted” as used herein, refers to independent replacementof one, two, three or more of the hydrogen atoms thereon withsubstituents including, but not limited to, deuterium, tritium, —F, —Cl,—Br, —I, —OH, C₁-C₁₂-alkyl; C₂-C₁₂-alkenyl, C₂-C₁₂-alkynyl, protectedhydroxy, —NO₂, —CN, —NH₂, —N₃, protected amino, alkoxy, thioalkoxy, oxo,thioxo, alkyl, -halo-C₂-C₁₂-alkenyl, -halo-C₂-C₁₂-alkynyl,-halo-C₃-C₁₂-cycloalkyl, —NH—C₂-C₁₂-alkenyl, —NH—C₂-C₁₂-alkynyl,—NH—C₃-C₁₂-cycloalkyl, —NH-aryl, —NH-heteroaryl, —NH-heterocycloalkyl,-dialkylamino, -diarylamino, -diheteroarylamino, —O—C₁-C₁₂-alkyl,—O—C₂-C₁₂-alkenyl, —O—C₂-C₁₂-alkynyl, —O—C₃-C₁₂-cycloalkyl, —O-aryl,—O-heteroaryl, —O— heterocycloalkyl, —C(O)—C₁-C₁₂-alkyl,—C(O)—C₂-C₁₂-alkenyl, —C(O)—C₂-C₁₂-alkynyl, —C(O)—C₃-C₁₂-cycloalkyl,—C(O)-aryl, —C(O)-heteroaryl, —C(O)-heterocycloalkyl, —CONH₂,—CONH—C₁-C₁₂-alkyl, —CONH—C₂-C₁₂-alkenyl, —CONH—C₂-C₁₂-alkynyl,—CONH—C₃-C₁₂-cycloalkyl, —CONH-aryl, —CONH-heteroaryl,—CONH-heterocycloalkyl, —OCO₂—C₁-C₁₂-alkyl, —OCO₂—C₂-C₁₂-alkenyl,—OCO₂—C₂-C₁₂-alkynyl, —OCO₂—C₃-C₁₂-cycloalkyl, —OCO₂-aryl,—OCO₂-heteroaryl, —OCO₂-heterocycloalkyl, —OCONH₂, —OCONH—C₁-C₁₂-alkyl,—OCONH—C₂-C₁₂-alkenyl, —OCONH—C₂-C₁₂-alkynyl, —OCONH—C₃-C₁₂-cycloalkyl,—OCONH-aryl, —OCONH-heteroaryl, —OCONH— heterocycloalkyl,—NHC(O)—C₁-C₁₂-alkyl, —NHC(O)—C₂-C₁₂-alkenyl, —NHC(O)—C₂-C₁₂-alkynyl,—NHC(O)—C₃-C₁₂-cycloalkyl, —NHC(O)-aryl, —NHC(O)-heteroaryl,—NHC(O)-heterocycloalkyl, —NHCO₂—C₁-C₁₂-alkyl, —NHCO₂—C₂-C₁₂-alkenyl,—NHCO₂—C₂-C₁₂-alkynyl, —NHCO₂—C₃-C₁₂-cycloalkyl, —NHCO₂-aryl,—NHCO₂-heteroaryl, —NHCO₂-heterocycloalkyl, —NHC(O)NH₂,—NHC(O)NH—C₁-C₁₂-alkyl, —NHC(O)NH—C₂-C₁₂-alkenyl,—NHC(O)NH—C₂-C₁₂-alkynyl, —NHC(O)NH—C₃-C₁₂-cycloalkyl, —NHC(O)NH-aryl,—NHC(O)NH-heteroaryl, —NHC(O)NH— heterocycloalkyl, NHC(S)NH₂,—NHC(S)NH—C₁-C₁₂-alkyl, —NHC(S)NH—C₂-C₁₂-alkenyl,—NHC(S)NH—C₂-C₁₂-alkynyl, —NHC(S)NH—C₃-C₁₂-cycloalkyl, —NHC(S)NH-aryl,—NHC(S)NH— heteroaryl, —NHC(S)NH-heterocycloalkyl, —NHC(NH)NH₂,—NHC(NH)NH—C₁-C₁₂-alkyl, —NHC(NH)NH—C₂-C₁₂-alkenyl,—NHC(NH)NH—C₂-C₁₂-alkynyl, —NHC(NH)NH—C₃-C₁₂-cycloalkyl,—NHC(NH)NH-aryl, —NHC(NH)NH-heteroaryl, —NHC(NH)NH-heterocycloalkyl,—NHC(NH)—C₁-C₁₂-alkyl, —NHC(NH)—C₂-C₁₂-alkenyl, —NHC(NH)—C₂-C₁₂-alkynyl,—NHC(NH)—C₃-C₁₂-cycloalkyl, —NHC(NH)-aryl, —NHC(NH)-heteroaryl,—NHC(NH)-heterocycloalkyl, —C(NH)NH—C₁-C₁₂-alkyl,—C(NH)NH—C₂-C₁₂-alkenyl, —C(NH)NH—C₂-C₁₂-alkynyl,—C(NH)NH—C₃-C₁₂-cycloalkyl, —C(NH)NH-aryl, —C(NH)NH-heteroaryl,—C(NH)NH-heterocycloalkyl, —S(O)—C₁-C₁₂-alkyl, —S(O)—C₂-C₁₂-alkenyl,—S(O)—C₂-C₁₂-alkynyl, —S(O)—C₃-C₁₂-cycloalkyl, —S(O)-aryl,—S(O)-heteroaryl, —S(O)-heterocycloalkyl —SO₂NH₂, —SO₂NH— C₁-C₁₂-alkyl,—SO₂NH— C₂-C₁₂-alkenyl, —SO₂NH— C₂-C₁₂-alkynyl, —SO₂NH—C₃-C₁₂-cycloalkyl, —SO₂NH— aryl, —SO₂NH-heteroaryl, —SO₂NH—heterocycloalkyl, —NHSO₂—C₁-C₁₂-alkyl, —NHSO₂—C₂-C₁₂-alkenyl,—NHSO₂—C₂-C₁₂-alkynyl, —NHSO₂—C₃-C₁₂-cycloalkyl, —NHSO₂-aryl,—NHSO₂-heteroaryl, —NHSO₂-heterocycloalkyl, —CH₂NH₂, —CH₂SO₂CH₃, -aryl,-arylalkyl, -heteroaryl, -heteroarylalkyl, -heterocycloalkyl,—C₃-C₁₂-cycloalkyl, polyalkoxyalkyl, polyalkoxy, -methoxymethoxy,-methoxyethoxy, —SH, —S—C₁-C₁₂-alkyl, —S—C₂-C₁₂-alkenyl,—S—C₂-C₁₂-alkynyl, —S—C₃-C₁₂-cycloalkyl, —S-aryl, —S-heteroaryl,—S-heterocycloalkyl, methylthiomethyl, or -L′-R′, wherein L′ isC₁-C₆alkylene, C₂-C₆alkenylene or C₂-C₆alkynylene, and R′ is aryl,heteroaryl, heterocyclic, C₃-C₁₂cycloalkyl or C₃-C₁₂cycloalkenyl. It isunderstood that the aryls, heteroaryls, alkyls, and the like can befurther substituted. In some cases, each substituent in a substitutedmoiety is additionally optionally substituted with one or more groups,each group being independently selected from C₁-C₆-alkyl, —F, —Cl, —Br,—I, —OH, —NO₂, —CN, and —NH₂.

The term “optionally substituted”, as used herein, means that thereferenced group may be substituted or unsubstituted. In one embodiment,the referenced group is optionally substituted with zero substituents,i.e., the referenced group is unsubstituted. In another embodiment, thereferenced group is optionally substituted with one or more additionalgroup(s) individually and independently selected from groups describedherein.

In accordance with the invention, any of the aryls, substituted aryls,heteroaryls and substituted heteroaryls described herein, can be anyaromatic group. Aromatic groups can be substituted or unsubstituted.

It is understood that any alkyl, alkenyl, alkynyl, cycloalkyl,heterocyclic and cycloalkenyl moiety described herein can also be analiphatic group or an alicyclic group.

An “aliphatic” group is a non-aromatic moiety comprised of anycombination of carbon atoms, hydrogen atoms, halogen atoms, oxygen,nitrogen or other atoms, and optionally contains one or more units ofunsaturation, e.g., double and/or triple bonds. Examples of aliphaticgroups are functional groups, such as alkyl, alkenyl, alkynyl, O, OH,NH, NH₂, C(O), S(O)₂, C(O)O, C(O)NH, OC(O)O, OC(O)NH, OC(O)NH₂, S(O)₂NH,S(O)₂NH₂, NHC(O)NH₂, NHC(O)C(O)NH, NHS(O)₂NH, NHS(O)₂NH₂, C(O)NHS(O)₂,C(O)NHS(O)₂NH or C(O)NHS(O)₂NH₂, and the like, groups comprising one ormore functional groups, non-aromatic hydrocarbons (optionallysubstituted), and groups wherein one or more carbons of a non-aromatichydrocarbon (optionally substituted) is replaced by a functional group.Carbon atoms of an aliphatic group can be optionally oxo-substituted. Analiphatic group may be straight chained, branched, cyclic, or acombination thereof and preferably contains between about 1 and about 24carbon atoms, more typically between about 1 and about 12 carbon atoms.In addition to aliphatic hydrocarbon groups, as used herein, aliphaticgroups expressly include, for example, alkoxyalkyls, polyalkoxyalkyls,such as polyalkylene glycols, polyamines, and polyimines, for example.Aliphatic groups may be optionally substituted.

The term “alicyclic,” as used herein, denotes a monovalent group derivedfrom a monocyclic or polycyclic saturated carbocyclic ring compound bythe removal of a single hydrogen atom. Examples include, but are notlimited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, bicyclo[2.2.1] heptyl, and bicyclo [2.2.2] octyl. Such alicyclic groups may befurther substituted.

It is understood that any alkyl, alkenyl, alkynyl, alicyclic,cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocyclic, aliphaticmoiety or the like, described herein can also be a divalent ormultivalent group when used as a linkage to connect two or more groupsor substituents, which can be at the same or different atom(s). One ofskill in the art can readily determine the valence of any such groupfrom the context in which it occurs.

The term “hydroxy activating group”, as used herein, refers to a labilechemical moiety which is known in the art to activate a hydroxy group sothat it will depart during synthetic procedures such as in asubstitution or elimination reactions. Examples of hydroxy activatinggroup include, but not limited to, mesylate, tosylate, triflate,p-nitrobenzoate, phosphonate and the like.

The term “activated hydroxy”, as used herein, refers to a hydroxy groupactivated with a hydroxy activating group, as defined above, includingmesylate, tosylate, triflate, p-nitrobenzoate, phosphonate groups, forexample.

The term “protected hydroxy,” as used herein, refers to a hydroxy groupprotected with a hydroxy protecting group, as defined above, includingbenzoyl, acetyl, trimethylsilyl, triethylsilyl, methoxymethyl groups,for example.

The term “hydroxy protecting group,” as used herein, refers to a labilechemical moiety which is known in the art to protect a hydroxy groupagainst undesired reactions during synthetic procedures. After saidsynthetic procedure(s) the hydroxy protecting group as described hereinmay be selectively removed. Hydroxy protecting groups as known in theare described generally in T. H. Greene and P. G., S. M. Wuts,Protective Groups in Organic Synthesis, 3rd edition, John Wiley & Sons,New York (1999). Examples of hydroxy protecting groups includebenzyloxycarbonyl, 4-nitrobenzyloxycarbonyl, 4-bromobenzyloxycarbonyl,4-methoxybenzyloxycarbonyl, methoxycarbonyl, tert-butoxycarbonyl,isopropoxycarbonyl, diphenylmethoxycarbonyl,2,2,2-trichloroethoxycarbonyl, 2-(trimethylsilyl)ethoxycarbonyl,2-furfuryloxycarbonyl, allyloxycarbonyl, acetyl, formyl, chloroacetyl,trifluoroacetyl, methoxyacetyl, phenoxyacetyl, benzoyl, methyl, t-butyl,2,2,2-trichloroethyl, 2-trimethylsilyl ethyl, 1,1-dimethyl-2-propenyl,3-methyl-3-butenyl, allyl, benzyl, para-methoxybenzyldiphenylmethyl,triphenylmethyl (trityl), tetrahydrofuryl, methoxymethyl,methylthiomethyl, benzyloxymethyl, 2,2,2-triehloroethoxymethyl,2-(trimethylsilyl)ethoxymethyl, methanesulfonyl, para-toluenesulfonyl,trimethylsilyl, triethylsilyl, triisopropylsilyl, and the like.Preferred hydroxy protecting groups for the present invention are acetyl(Ac or —C(O)CH₃), benzoyl (Bz or —C(O)C₆H₅), and trimethylsilyl (TMS or—Si(CH₃)₃), and the like.

The term “hydroxy prodrug group,” as used herein, refers to a promoietygroup which is known in the art to change the physicochemical, and hencethe biological properties of a parent drug in a transient manner bycovering or masking the hydroxy group. After said syntheticprocedure(s), the hydroxy prodrug group as described herein must becapable of reverting back to hydroxy group in vivo. Hydroxy prodruggroups as known in the art are described generally in Kenneth B. Sloan,Prodrugs, Topical and Ocular Drug Delivery, (Drugs and thePharmaceutical Sciences; Volume 53), Marcel Dekker, Inc., New York(1992).

The term “amino protecting group,” as used herein, refers to a labilechemical moiety which is known in the art to protect an amino groupagainst undesired reactions during synthetic procedures. After saidsynthetic procedure(s) the amino protecting group as described hereinmay be selectively removed. Amino protecting groups as known in the artare described generally in T. H. Greene and P. G. M. Wuts, ProtectiveGroups in Organic Synthesis, 3rd edition, John Wiley & Sons, New York(1999). Examples of amino protecting groups include, but are not limitedto, methoxycarbonyl, t-butoxycarbonyl, 9-fluorenyl-methoxycarbonyl,benzyloxycarbonyl, and the like.

The term “protected amino,” as used herein, refers to an amino groupprotected with an amino protecting group as defined above.

The term “leaving group” means a functional group or atom which can bedisplaced by another functional group or atom in a substitutionreaction, such as a nucleophilic substitution reaction. By way ofexample, representative leaving groups include chloro, bromo and iodogroups; sulfonic ester groups, such as mesylate, tosylate, brosylate,nosylate and the like; and acyloxy groups, such as acetoxy,trifluoroacetoxy and the like.

The compounds described herein contain one or more asymmetric centersand thus give rise to enantiomers, diastereomers, and otherstereoisomeric forms that may be defined, in terms of absolutestereochemistry, as (R)- or (S), or as (D)- or (L)- for amino acids. Thepresent invention is meant to include all such possible isomers, as wellas their racemic and optically pure forms. Optical isomers may beprepared from their respective optically active precursors by theprocedures described above, or by resolving the racemic mixtures. Theresolution can be carried out in the presence of a resolving agent, bychromatography or by repeated crystallization or by some combination ofthese techniques, which are known to those skilled in the art. Furtherdetails regarding resolutions can be found in Jacques, et al.,Enantiomers, Racemates, and Resolutions (John Wiley & Sons, 1981). Whenthe compounds described herein contain olefinic double bonds or othercenters of geometric asymmetry, and unless specified otherwise, it isintended that the compounds include both E and Z geometric isomers.Likewise, all tautomeric forms are also intended to be included. Theconfiguration of any carbon-carbon double bond appearing herein isselected for convenience only and is not intended to designate aparticular configuration unless the text so states; thus a carbon-carbondouble bond depicted arbitrarily herein as trans may be cis, trans, or amixture of the two in any proportion.

In certain embodiments, the compounds of each formula herein are definedto include isotopically labelled compounds. An “isotopically labelledcompound” is a compound in which at least one atomic position isenriched in a specific isotope of the designated element to a levelwhich is significantly greater than the natural abundance of thatisotope. For example, one or more hydrogen atom positions in a compoundcan be enriched with deuterium to a level which is significantly greaterthan the natural abundance of deuterium, for example, enrichment to alevel of at least 1%, preferably at least 20% or at least 50%. Such adeuterated compound may, for example, be metabolized more slowly thanits non-deuterated analog, and therefore exhibit a longer half-life whenadministered to a subject. Such compounds can synthesize using methodsknown in the art, for example by employing deuterated startingmaterials. Unless stated to the contrary, isotopically labelledcompounds are pharmaceutically acceptable.

Certain compounds of the present invention may also exist in differentstable conformational forms which may be separable. Torsional asymmetrydue to restricted rotation about an asymmetric single bond, for examplebecause of steric hindrance or ring strain, may permit separation ofdifferent conformers. The present invention includes each conformationalisomer of these compounds and mixtures thereof.

The term “subject” as used herein refers to a mammal. A subjecttherefore refers to, for example, dogs, cats, horses, cows, pigs, guineapigs, and the like. Preferably the subject is a human. When the subjectis a human, the subject may be referred to herein as a patient.

As used herein, the term “pharmaceutically acceptable salt” refers tothose salts of the compounds formed by the process of the presentinvention which are, within the scope of sound medical judgment,suitable for use in contact with the tissues of humans and lower animalswithout undue toxicity, irritation, allergic response and the like, andare commensurate with a reasonable benefit/risk ratio. Pharmaceuticallyacceptable salts are well known in the art.

Berge, et al. describes pharmaceutically acceptable salts in detail inJ. Pharmaceutical Sciences, 66: 1-19 (1977). The salts can be preparedin situ during the final isolation and purification of the compounds ofthe invention, or separately by reacting the free base function with asuitable organic acid. Examples of pharmaceutically acceptable saltsinclude, but are not limited to, nontoxic acid addition salts e.g.,salts of an amino group formed with inorganic acids such as hydrochloricacid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloricacid or with organic acids such as acetic acid, maleic acid, tartaricacid, citric acid, succinic acid or malonic acid or by using othermethods used in the art such as ion exchange. Other pharmaceuticallyacceptable salts include, but are not limited to, adipate, alginate,ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate,butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate,digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate,glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate,hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate,lactate, laurate, lauryl sulfate, malate, maleate, malonate,methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate,oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate,phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate,tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts,and the like. Representative alkali or alkaline earth metal saltsinclude sodium, lithium, potassium, calcium, magnesium, and the like.Further pharmaceutically acceptable salts include, when appropriate,nontoxic ammonium, quaternary ammonium, and amine cations formed usingcounterions such as halide, hydroxide, carboxylate, sulfate, phosphate,nitrate, alkyl having from 1 to 6 carbon atoms, sulfonate and arylsulfonate.

Pharmaceutically acceptable salts can also be prepared by deprotonationof the parent compound with a suitable base, thereby forming the anionicconjugate base of the parent compound. In such salts the counter ion isa cation. Suitable cations include ammonium and metal cations, such asalkali metal cations, including Li⁺, Na⁺, K⁺ and Cs⁺, and alkaline earthmetal cations, such as Mg²⁺ and Ca²⁺.

As used herein, the term “pharmaceutically acceptable ester” refers toesters of the compounds formed by the process of the present inventionwhich hydrolyze in vivo and include those that break down readily in thehuman body to leave the parent compound or a salt thereof. Suitableester groups include, for example, those derived from pharmaceuticallyacceptable aliphatic carboxylic acids, particularly alkanoic, alkenoic,cycloalkanoic and alkanedioic acids, in which each alkyl or alkenylmoiety advantageously has not more than 6 carbon atoms. Examples ofparticular esters include, but are not limited to, formates, acetates,propionates, butyrates, acrylates and ethylsuccinates.

The term “pharmaceutically acceptable prodrugs” as used herein refers tothose prodrugs of the compounds formed by the process of the presentinvention which are, within the scope of sound medical judgment,suitable for use in contact with the tissues of humans and lower animalswith undue toxicity, irritation, allergic response, and the like,commensurate with a reasonable benefit/risk ratio, and effective fortheir intended use, as well as the zwitterionic forms, where possible,of the compounds of the present invention. “Prodrug”, as used hereinmeans a compound, which is convertible in vivo by metabolic means (e.g.by hydrolysis) to afford any compound delineated by the formulae of theinstant invention. Various forms of prodrugs are known in the art, forexample, as discussed in Bundgaard, (ed.), Design of Prodrugs, Elsevier(1985); Widder, et al. (ed.), Methods in Enzymology, Vol. 4, AcademicPress (1985); Krogsgaard-Larsen, et al., (ed.). “Design and Applicationof Prodrugs, Textbook of Drug Design and Development, Chapter 5, 113-191(1991); Bundgaard, et al., Journal of Drug Deliver Reviews, 8:1-38(1992); Bundgaard, J. of Pharmaceutical Sciences, 77:285 et seq. (1988);Higuchi and Stella (eds.) Prodrugs as Novel Drug Delivery Systems,American Chemical Society (1975); and Bernard Testa & Joachim Mayer,“Hydrolysis In Drug And Prodrug Metabolism: Chemistry, Biochemistry AndEnzymology,” John Wiley and Sons, Ltd. (2002).

Additional types of prodrugs are also encompassed. For instance, freecarboxyl groups can be derivatized as amides or alkyl esters. Freehydroxy groups may be derivatized using groups including but not limitedto hemisuccinates, ethyl succinate, phosphate esters,dimethylaminoacetates, and phosphoryloxymethyloxycarbonyls, as outlinedin Advanced Drug Delivery Reviews, 1996, 19, 115. Carbamate prodrugs ofhydroxy and amino groups are also included, as are carbonate prodrugs,sulfonate esters and sulfate esters of hydroxy groups. Derivatization ofhydroxy groups as (acyloxy)methyl and (acyloxy)ethyl ethers wherein theacyl group may be an alkyl ester, optionally substituted with groupsincluding but not limited to ether, amine and carboxylic acidfunctionalities, or where the acyl group is an amino acid ester asdescribed above, are also encompassed. Prodrugs of this type aredescribed in J. Med. Chem. 1996, 39, 10. Free amines can also bederivatized as amides, sulfonamides or phosphonamides. All of theseprodrug moieties may incorporate groups including but not limited toether, amine and carboxylic acid functionalities. In certainembodiments, a compound of the invention can incorporate two or moregroups that are metabolically removed in vivo to yield the active parentcompound. For example, a compound of formula I wherein R₁ is an aminoacid residue can also be esterified, for example at a hydroxyl group ofthe sugar residue, to form a compound with two groups that can beremoved in vivo to yield the active compound.

The term “treating”, as used herein, means relieving, lessening,reducing, eliminating, modulating, or ameliorating, i.e. causingregression of the disease state or condition. Treating can also includeinhibiting, i.e. arresting the development, of a existing disease stateor condition, and relieving or ameliorating, i.e. causing regression ofan existing disease state or condition, for example when the diseasestate or condition may already be present.

The term “preventing”, as used herein means, to completely or almostcompletely stop a disease state or condition, from occurring in apatient or subject, especially when the patient or subject ispredisposed to such or at risk of contracting a disease state orcondition.

Additionally, the compounds of the present invention, for example, thesalts of the compounds, can exist in either hydrated or unhydrated (theanhydrous) form or as solvates with other solvent molecules. Nonlimitingexamples of hydrates include monohydrates, dihydrates, etc. Nonlimitingexamples of solvates include ethanol solvates, acetone solvates, etc.

“Solvates” means solvent addition forms that contain eitherstoichiometric or non stoichiometric amounts of solvent. Some compoundshave a tendency to trap a fixed molar ratio of solvent molecules in thecrystalline solid state, thus forming a solvate. If the solvent is waterthe solvate formed is a hydrate, when the solvent is alcohol, thesolvate formed is an alcoholate. Hydrates are formed by the combinationof one or more molecules of water with one of the substances in whichthe water retains its molecular state as H₂O, such combination beingable to form one or more hydrates.

As used herein, the term “analog” refers to a chemical compound that isstructurally similar to another but differs slightly in composition (asin the replacement of one atom by an atom of a different element or inthe presence of a particular functional group, or the replacement of onefunctional group by another functional group). Thus, an analog is acompound that is similar to or comparable in function and appearance tothe reference compound.

The term “aprotic solvent,” as used herein, refers to a solvent that isrelatively inert to proton activity, i.e., not acting as a proton-donor.Examples include, but are not limited to, hydrocarbons, such as hexaneand toluene, for example, halogenated hydrocarbons, such as, forexample, methylene chloride, ethylene chloride, chloroform, and thelike, heterocyclic compounds, such as, for example, tetrahydrofuran andN-methylpyrrolidinone, and ethers such as diethyl ether,bis-methoxymethyl ether. Such solvents are well known to those skilledin the art, and individual solvents or mixtures thereof may be preferredfor specific compounds and reaction conditions, depending upon suchfactors as the solubility of reagents, reactivity of reagents andpreferred temperature ranges, for example. Further discussions ofaprotic solvents may be found in organic chemistry textbooks or inspecialized monographs, for example: Organic Solvents PhysicalProperties and Methods of Purification, 4th ed., edited by John A.Riddick et al., Vol. II, in the Techniques of Chemistry Series, JohnWiley & Sons, N Y, 1986.

The terms “protogenic organic solvent” or “protic solvent” as usedherein, refer to a solvent that tends to provide protons, such as analcohol, for example, methanol, ethanol, propanol, isopropanol, butanol,t-butanol, and the like. Such solvents are well known to those skilledin the art, and individual solvents or mixtures thereof may be preferredfor specific compounds and reaction conditions, depending upon suchfactors as the solubility of reagents, reactivity of reagents andpreferred temperature ranges, for example. Further discussions ofprotogenic solvents may be found in organic chemistry textbooks or inspecialized monographs, for example: Organic Solvents PhysicalProperties and Methods of Purification, 4th ed., edited by John A.Riddick et al., Vol. II, in the Techniques of Chemistry Series, JohnWiley & Sons, N Y, 1986.

Combinations of substituents and variables envisioned by this inventionare only those that result in the formation of stable compounds. Theterm “stable”, as used herein, refers to compounds which possessstability sufficient to allow manufacture and which maintains theintegrity of the compound for a sufficient period of time to be usefulfor the purposes detailed herein (e.g., therapeutic or prophylacticadministration to a subject).

The synthesized compounds can be separated from a reaction mixture andfurther purified by a method such as column chromatography, highpressure liquid chromatography, or recrystallization. Additionally, thevarious synthetic steps may be performed in an alternate sequence ororder to give the desired compounds. In addition, the solvents,temperatures, reaction durations, etc. delineated herein are forpurposes of illustration only and variation of the reaction conditionscan produce the desired bridged macrocyclic products of the presentinvention. Synthetic chemistry transformations and protecting groupmethodologies (protection and deprotection) useful in synthesizing thecompounds described herein include, for example, those described in R.Larock, Comprehensive Organic Transformations, VCH Publishers (1989); T.W. Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis, 2d.Ed., John Wiley and Sons (1991); L. Fieser and M. Fieser, Fieser andFieser's Reagents for Organic Synthesis, John Wiley and Sons (1994); andL. Paquette, ed., Encyclopedia of Reagents for Organic Synthesis, JohnWiley and Sons (1995).

The compounds of this invention may be modified by appending variousfunctionalities via synthetic means delineated herein to enhanceselective biological properties. Such modifications include those whichincrease biological penetration into a given biological system (e.g.,blood, lymphatic system, central nervous system), increase oralavailability, increase solubility to allow administration by injection,alter metabolism and alter rate of excretion.

Pharmaceutical Compositions

The pharmaceutical compositions of the present invention comprise atherapeutically effective amount of a compound of the present inventionformulated together with one or more pharmaceutically acceptablecarriers. As used herein, the term “pharmaceutically acceptable carrier”means a non-toxic, inert solid, semi-solid or liquid filler, diluent,encapsulating material or formulation auxiliary of any type. Someexamples of materials which can serve as pharmaceutically acceptablecarriers are sugars such as lactose, glucose and sucrose; starches suchas corn starch and potato starch; cellulose and its derivatives such assodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate;powdered tragacanth; malt; gelatin; talc; excipients such as cocoabutter and suppository waxes; oils such as peanut oil, cottonseed oil;safflower oil; sesame oil; olive oil; corn oil and soybean oil; glycols;such a propylene glycol; esters such as ethyl oleate and ethyl laurate;agar; buffering agents such as magnesium hydroxide and aluminumhydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer'ssolution; ethyl alcohol, and phosphate buffer solutions, as well asother non-toxic compatible lubricants such as sodium lauryl sulfate andmagnesium stearate, as well as coloring agents, releasing agents,coating agents, sweetening, flavoring and perfuming agents,preservatives and antioxidants can also be present in the composition,according to the judgment of the formulator. The pharmaceuticalcompositions of this invention can be administered to humans and otheranimals orally, rectally, parenterally, intracisternally,intravaginally, intraperitoneally, topically (as by powders, ointments,or drops), buccally, or as an oral or nasal spray.

The pharmaceutical compositions of this invention may be administeredorally, parenterally, by inhalation spray, topically, rectally, nasally,buccally, vaginally or via an implanted reservoir, preferably by oraladministration or administration by injection. The pharmaceuticalcompositions of this invention may contain any conventional non-toxicpharmaceutically-acceptable carriers, adjuvants or vehicles. In somecases, the pH of the formulation may be adjusted with pharmaceuticallyacceptable acids, bases or buffers to enhance the stability of theformulated compound or its delivery form. The term parenteral as usedherein includes subcutaneous, intracutaneous, intravenous,intramuscular, intraarticular, intraarterial, intrasynovial,intrasternal, intrathecal, intralesional and intracranial injection orinfusion techniques.

Liquid dosage forms for oral administration include pharmaceuticallyacceptable emulsions, microemulsions, solutions, suspensions, syrups andelixirs. In addition to the active compounds, the liquid dosage formsmay contain inert diluents commonly used in the art such as, forexample, water or other solvents, solubilizing agents and emulsifierssuch as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethylacetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyleneglycol, dimethylformamide, oils (in particular, cottonseed, groundnut,corn, germ, olive, castor, and sesame oils), glycerol,tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid estersof sorbitan, and mixtures thereof. Besides inert diluents, the oralcompositions can also include adjuvants such as wetting agents,emulsifying and suspending agents, sweetening, flavoring, and perfumingagents.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions may be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectablesolution, suspension or emulsion in a nontoxic parenterally acceptablediluent or solvent, for example, as a solution in 1,3-butanediol. Amongthe acceptable vehicles and solvents that may be employed are water,Ringer's solution, U.S.P. and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables.

The injectable formulations can be sterilized, for example, byfiltration through a bacterial-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved or dispersed in sterile water or other sterile injectablemedium prior to use.

In order to prolong the effect of a drug, it is often desirable to slowthe absorption of the drug from subcutaneous or intramuscular injection.This may be accomplished by the use of a liquid suspension ofcrystalline or amorphous material with poor water solubility. The rateof absorption of the drug then depends upon its rate of dissolution,which, in turn, may depend upon crystal size and crystalline form.Alternatively, delayed absorption of a parenterally administered drugform is accomplished by dissolving or suspending the drug in an oilvehicle. Injectable depot forms are made by forming microencapsulematrices of the drug in biodegradable polymers such aspolylactide-polyglycolide. Depending upon the ratio of drug to polymerand the nature of the particular polymer employed, the rate of drugrelease can be controlled. Examples of other biodegradable polymersinclude poly(orthoesters) and poly(anhydrides). Depot injectableformulations are also prepared by entrapping the drug in liposomes ormicroemulsions which are compatible with body tissues.

Compositions for rectal or vaginal administration are preferablysuppositories which can be prepared by mixing the compounds of thisinvention with suitable non-irritating excipients or carriers such ascocoa butter, polyethylene glycol or a suppository wax which are solidat ambient temperature but liquid at body temperature and therefore meltin the rectum or vaginal cavity and release the active compound.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activecompound is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or: a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid, b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia, c) humectants such as glycerol, d) disintegratingagents such as agar-agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate, e) solutionretarding agents such as paraffin, f) absorption accelerators such asquaternary ammonium compounds, g) wetting agents such as, for example,cetyl alcohol and glycerol monostearate, h) absorbents such as kaolinand bentonite clay, and i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof. In the case of capsules, tablets and pills, thedosage form may also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polyethylene glycols andthe like.

The active compounds can also be in micro-encapsulated form with one ormore excipients as noted above. The solid dosage forms of tablets,dragees, capsules, pills, and granules can be prepared with coatings andshells such as enteric coatings, release controlling coatings and othercoatings well known in the pharmaceutical formulating art. In such soliddosage forms the active compound may be admixed with at least one inertdiluent such as sucrose, lactose or starch. Such dosage forms may alsocomprise, as is normal practice, additional substances other than inertdiluents, e.g., tableting lubricants and other tableting aids such amagnesium stearate and microcrystalline cellulose. In the case ofcapsules, tablets and pills, the dosage forms may also comprisebuffering agents. They may optionally contain opacifying agents and canalso be of a composition that they release the active ingredient(s)only, or preferentially, in a certain part of the intestinal tract,optionally, in a delayed manner. Examples of embedding compositionswhich can be used include polymeric substances and waxes.

Dosage forms for topical or transdermal administration of a compound ofthis invention include ointments, pastes, creams, lotions, gels,powders, solutions, sprays, inhalants or patches. The active componentis admixed under sterile conditions with a pharmaceutically acceptablecarrier and any needed preservatives or buffers as may be required.Ophthalmic formulation, ear drops, eye ointments, powders and solutionsare also contemplated as being within the scope of this invention.

The ointments, pastes, creams and gels may contain, in addition to anactive compound of this invention, excipients such as animal andvegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulosederivatives, polyethylene glycols, silicones, bentonites, silicic acid,talc and zinc oxide, or mixtures thereof.

Powders and sprays can contain, in addition to the compounds of thisinvention, excipients such as lactose, talc, silicic acid, aluminumhydroxide, calcium silicates and polyamide powder, or mixtures of thesesubstances. Sprays can additionally contain customary propellants suchas chlorofluorohydrocarbons.

Transdermal patches have the added advantage of providing controlleddelivery of a compound to the body. Such dosage forms can be made bydissolving or dispensing the compound in the proper medium. Absorptionenhancers can also be used to increase the flux of the compound acrossthe skin. The rate can be controlled by either providing a ratecontrolling membrane or by dispersing the compound in a polymer matrixor gel.

Unless otherwise defined, all technical and scientific terms used hereinare accorded the meaning commonly known to one with ordinary skill inthe art. All publications, patents, published patent applications, andother references mentioned herein are hereby incorporated by referencein their entirety.

Abbreviations

Abbreviations which have been used in the descriptions of the schemesand the examples that follow are:

ACN for acetonitrile;

BME for 2-mercaptoethanol;

BOP for benzotriazol-1-yloxy-tris(dimethylamino)phosphoniumhexafluorophosphate;

BTC for bis(trichloromethyl)carbonate; triphosgene;

BzCl for benzoyl chloride;

CAN for ceric ammonium nitrate;

CDI for carbonyldiimidazole;

COD for cyclooctadiene;

DABCO for 1,4-diazabicyclo[2.2.2]octane;

DAST for diethylaminosulfur trifluoride;

DABCYL for 6-(N-4′-carboxy-4-(dimethylamino)azobenzene)-aminohexyl-;

1-O-(2-cyanoethyl)-(N,N-diisopropyl)-phosphoramidite;

DBU for 1,8-Diazabicycloundec-7-ene;

DCC for N,N′-dicyclohexylcarbodiimide;

DCM for dichloromethane;

DIAD for diisopropyl azodicarboxylate;

DIBAL-H for diisobutylaluminum hydride;

DIPEA for diisopropyl ethylamine;

DMAP for N,N-dimethylaminopyridine;

DME for ethylene glycol dimethyl ether;

DMEM for Dulbecco's Modified Eagles Media;

DMF for N,N-dimethyl formamide;

DMSO for dimethylsulfoxide;

DSC for N,N′-disuccinimidyl carbonate;

DUPHOS for

EDANS for 5-(2-Amino-ethylamino)-naphthalene-1-sulfonic acid;EDCI or EDC for 1-(3-diethylaminopropyl)-3-ethylcarbodiimidehydrochloride;EtOAc for ethyl acetate;EtOH for ethyl alcohol;HATU for O (7-Azabenzotriazole-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate;HCl for hydrochloric acid;Hoveyda's Cat. for Dichloro(o-isopropoxyphenylmethylene)(tricyclohexylphosphine)ruthenium(II);In for indium;KHMDS is potassium bis(trimethylsilyl) amide;Ms for mesyl;NMM for N-4-methylmorpholine;NMI for N-methylimidazole;NMO for N-4-methylmorpholine-N-Oxide;PMB for para-methoxybenzyl;PyBrOP for Bromo-tri-pyrolidino-phosphonium hexafluorophosphate;Ph for phenyl;RCM for ring-closing metathesis;RT for reverse transcription;RT-PCR for reverse transcription-polymerase chain reaction;TBME for tert-butyl methyl ether;TCDI for 1,1′-thiocarbonyldiimidazole;TEA for triethyl amine;Tf₂O for trifluoromethanesulfonic anhydride;TFA for trifluoroacetic acid;THF for tetrahydrofuran;TLC for thin layer chromatography;(TMS)₂NH for hexamethyldisilazane;TMSOTf for trimethylsilyl trifluoromethanesulfonate;TBS for t-Butyldimethylsilyl;TMS for trimethylsilyl;TPAP tetrapropylammonium perruthenate;TPP or PPh₃ for triphenylphosphine;TrCl for trityl chloride;DMTrCl for 4,4′-dimethoxytrityl chloride;tBOC or Boc for tert-butyloxy carbonyl;Xantphos for 4,5-Bis-diphenylphosphanyl-9,9-dimethyl-9H-xanthene;X-Phos for 2-(dicyclohexylphosphino)-2′,4′,6′-triisopropylbiphenyl; andZhan 1 B for

Synthetic Methods

The compounds and processes of the present invention will be betterunderstood in connection with the following synthetic schemes thatillustrate the methods by which the compounds of the invention may beprepared, which are intended as an illustration only and not to limitthe scope of the invention. Various changes and modifications to thedisclosed embodiments will be apparent to those skilled in the art andsuch changes and modifications including, without limitation, thoserelating to the chemical structures, substituents, derivatives, and/ormethods of the invention may be made without departing from the spiritof the invention and the scope of the appended claims.

As shown in Scheme 1-1, novel RSV analogs of the compound of formula 8are prepared starting from compounds 1 and 2. Compound 2 can beprotected by groups such as, but not limited to, tert-butyloxycarbonyl(BOC), acetyl (Ac) and 2-(trimethylsilyl-ethoxy)methyl (SEM). Initially,compounds 1 and 2 are heated to reflux in the presence of K₂CO₃ to formcompound 3. Compound 3 is converted to the corresponding aniline 4 usingreduction conditions such as, but not limited to, Fe/NH4Cl in hot EtOH.Compound 4 is cyclized using amide coupling conditions like, but notrestricted to, 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluroniumhexafluorophosphate (HBTU) and Et₃N in DMF at ambient temperature toyield compound 5. Compound 5 is reacted with an alkyl, aryl orheteroaryl halide to produce compound 6. The transformation requireseither C—N cross-coupling conditions using Pd or Cu catalysts oralkylation conditions requiring an inorganic or amine base. Deprotectionof compound 6 with appropriate reaction conditions such as, but notlimited to, acid, base or a nucleophilic fluoride source yields compound7. Compound 7 is reacted with either an aryl, heteroaryl or alkyl halidevia a displacement of the halogen (X) or via suitable couplingconditions using Pd or Cu catalysts to afford compound 8.

Scheme 1-2 illustrates alternative methods for assembling compoundshaving oxadiazole rings. Compounds 7 can be reacted with oxadiazolone 9in the presence of a coupling reagent such as, but not limited to,(benzotriazol-1-yloxy)tris(dimethylamino)phosphonium hexafluorophosphate(BOP) to afford compounds 10, containing an oxadiazole ring.Alternatively, compound 7 is reacted with 1,1′-carbonyldiimidazole (CDI)to yield the 1H-imidazole-1-carboxamide intermediate 11. Reaction ofthis intermediate and hydrazide 12 produces the amino semicarbazide 13,which is cyclized to the oxadiazole compound 10 using 4-toluenesulfonylchloride (TsCl), phosphorus(V) oxychloride (POCl₃) or other activatingreagents.

As shown in Scheme 1-3, two separate classes of compounds, 15 and 17,are made from 7. Compound 7 is reacted with alkyl, aryl or heteroarylisocyanate 14 to yield urea-based compounds 15. Alternatively, amine 7is reacted with acyl halide 16 to furnish amide compounds 17.

As shown in Scheme 2, amine intermediate 25 is prepared starting fromcompounds 18 and 3-ethoxy-3-oxopropanoic acid. Diamine 18 and3-ethoxy-3-oxopropanoic acid are coupled using, but not limited to,N,N′-dicyclohexylcarbodiimide (DCC), which yields intermediates 19. Thecarboxylic acids 20 are furnished through the saponification of 19.Compound 20 is cyclized to 21 using coupling reagents such as, but notlimited to, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI) and1-hydroxybenzotriazole (HOBt) or phosphorus pentachloride (PCl₅). ThePMB-protected amide 22 is furnished through reaction of the amide 21with K₂CO₃ and 4-methoxybenzyl chloride (PMBCl). The introduction of anazide to the malonamide derivative 22 is achieved through reaction withKOtBu and 2,4,6-triisopropylbenzenesulfonyl azide. The reduction ofazide 23 to amine 24 is accomplished using palladium on carbon under ahydrogen atmosphere. Amine 25 is furnished upon cleavage of the PMBgroup using ceric ammonium nitrate (CAN).

Illustrated in Scheme 3, novel RSV analogs of the compounds of formula26 and 28 are prepared starting from compound 25. Compound 25 is reactedwith either an aryl, heteroaryl or alkyl halide via a displacement ofthe halogen (X) or via suitable coupling conditions using Pd or Cucatalysts to afford compounds 26. Additionally, amine 25 is reacted with1,1′-thiocarbonyldiimidazole followed by hydrazide 12 to furnishthiosemicarbazide 27. Compound 28 is synthesized by cyclization of 27using, but not limited to, EDCI and DMF at elevated temperatures.

Scheme 4 depicts the various synthetic routes of hydrazide 31 startingfrom ester 29, wherein

is aryl, heteroaryl, —C₃-C₁₂ cycloalkyl, or 3- to 12-memberedheterocycloalkyl. Compound 29 is reacted with morpholine via adisplacement of the halogen (X) or via suitable coupling conditionsusing Pd or Cu catalysts to afford compound 30. The ester 30 isconverted to the hydrazide 31 upon subjection to hydrazine hydrate andEtOH at either ambient or elevated temperatures. Alternatively,hydrazide 31 can be made in reverse fashion where 29 is converted to thehydrazide 32, which to subsequently converted to 31 when reacted withmorpholine. Furthermore, 31 can be synthesized without the use ofhydrazine. Ester 29 is hydrolyzed using a hydroxide base such as LiOH.Next, the carboxylic acid 33 is reacted with BOC-protected hydrazine anda coupling reagents, such as, but not limited to,1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium3-oxid hexafluorophosphate (HATU), which yields intermediate 34.Subjection of the halide-containing compound 34 and morpholine todisplacement or coupling conditions furnishes 35, which uponacid-promoted deprotection of the BOC group, yields hydrazide 31.

EXAMPLES

The compounds and processes of the present invention will be betterunderstood in connection with the following examples, which are intendedas an illustration only and not limiting of the scope of the invention.Various changes and modifications to the disclosed embodiments will beapparent to those skilled in the art and such changes and modificationsincluding, without limitation, those relating to the chemicalstructures, substituents, derivatives, formulations and/or methods ofthe invention may be made without departing from the spirit of theinvention and the scope of the appended claims. Examples 1-18 wereprepared and tested as the indicated enantiomer. Examples 19-21 wereprepared and tested as a racemic mixture.

Example 1

Example 1 Step a

A mixture of 1-fluoro-2-nitrobenzene (4.23 g, 30 mmol), K₂CO₃ (12.42 g,90 mmol) and (S)-3-amino-2-(tert-butoxycarbonylamino)propanoic acid(6.73 g, 33 mmol) in EtOH (100 mL) were refluxed for 16 hours. Thereaction mixture was poured into water and adjusted to pH=3-4. Then itwas extracted with EtOAc. The organic layer was dried over Na₂SO₄,filtered and concentrated to afford(S)-2-((tert-butoxycarbonyl)amino-3-((2-nitrophenyl)amino)propanoic acidas a red solid (9.4 g) that was used without further purification.ESI-MS m/z: 326.50 [M+H]⁺.

Example 1 Step b

A solution of the compound from step a (9.4 g, 28.9 mmol), Fe (6.48 g,115.6 mmol) and saturated NH₄Cl (45 mL) in EtOH (200 mL) was stirred at80° C. for 1.5 hrs. Solid was filtered through celite and the filtratewas diluted with EtOAc, washed with water and separated. The organiclayer was dried over Na₂SO₄, filtered and concentrated under vacuum toafford (S)-3-((20aminophenyl)amino)-2-((tert-butoxycarbonyl)amino)propanoic acid as a dark solid (10.6 g) that was used without furtherpurification. ESI-MS m/z: 296.00 [M+H]⁺.

Example 1 Step c

A solution of the compound from step b (10.6 g, 36 mmol), HBTU (12.7 g,39.6 mmol) and Et₃N (10.9 g, 108 mmol) in DMF (200 mL) was stirred atr.t. for 2 hrs. The reaction mixture was diluted with EtOAc, washed withwater and brine and separated. The organic layer was dried over Na₂SO₄,filtered and concentrated. The crude product was purified by Flash(MeCN/H₂O) to give tert-butyl(S)-(2-oxo-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-3-yl)carbmate(5.0 g) as a yellow solid ESI-MS m/z: 278.05 [M+H]⁺.

Example 1 Step d

A mixture of the compound from step c (1.0 g, 36 mmol), bromobenzene(1.13 g, 7.22 mmol), X-Phos (343 mg, 0.72 mmol), Pd₂(dba)₃ (745 mg, 0.72mmol) and K₂CO₃ (2.48 g, 18 mmol) in tBuOH (20 mL) was degassed andstirred at 80° C. for 12 hrs under N₂. The crude product was purified bysilica gel column to give tert-butyl(S)-(4-oxo-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-3-yl)carbamate(240 mg) as a yellow solid (370 mg). ESI-MS m/z: 354.25 [M+H]⁺.

Example 1 Step e

A mixture of the compound from step d (370 mg) in DCM (5 mL) was addedTFA (2.5 mL). The reaction was stirred at r.t. for 2 hrs. Solvent wasremoved and the residue was basified with ammonia, extracted with DCM,washed with water and brine and separated. The organic layer was driedover Na₂SO₄, filtered and concentrated under vacuum to afford the titlecompound (240 mg) as a white solid ESI-MS m/z=254.20 [M+H]⁺.

Example 1 Step f

A solution of the compound from step e (50 mg, 0.2 mmol),5-phenyl-1,3,4-oxadiazol-2(3H)-one (65 mg, 0.4 mmol), BOP (220 mg, 0.5mmol) and DIPEA (104 mg, 0.8 mmol) in DMF (1 mL) was stirred at r.t. for48 hrs. The crude product was diluted with EtOAc, washed with water (×3)and brine. The organic layer was dried over Na₂SO₄, filtered andconcentrated to dryness. The crude mixture was purified by Prep-HPLC(MeCN/H₂O) to give(S)-5-phenyl-3-((5-phenyl-1,3,4-oxadiazol-2-yl)amino)-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-oneas a white solid (15.3 mg). ESI-MS m/z: 398.10 [M+H]⁺.

Example 2

A mixture of the compound from the step e of example 1 (50 mg, 0.2mmol), 2-chlorobenzo[d]oxazole (60 mg, 0.4 mmol), Et₃N (104 mg, 1.0mmol) in DMF (2 mL) was stirred at 60° C. for 12 hrs. The reactionmixture was diluted with EtOAc, washed with water (×3) and brine. Theorganic layer was dried over Na₂SO₄, filtered and concentrated todryness. The crude product was purified by Prep-HPLC (MeCN/H₂O) to give(S)-3-(benzo[d]oxazol-2-ylamino)-5-phenyl-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-oneas a white solid (26.6 mg). ESI-MS m/z: 371.05 [M+H]⁺.

Example 3

Example 3 was prepared from the compound (step e in example 1) andphenyl isocyanate. ESI-MS m/z: 373.05 [M+H]⁺.

Example 4

A mixture of the compound from the step e of example 1 (50 mg, 0.2mmol), benzoic acid (49 mg, 0.4 mmol), HATU (152 mg, 0.4 mmol) and DIPEA(52 mg, 0.4 mmol) in DMF (2 mL) was stirred at r.t. for 2 hrs. The crudeproduct was purified by Prep-HPLC (MeCN/H₂O) to give(S)—N-(4-oxo-1-phenyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-3-yl)benzamideas a white solid (34.4 mg). ESI-MS m/z: 358.10 [M+H]⁺. ¹H NMR (300 MHz,DMSO-d₆) δ 3.88-4.26 (m, 2H), 4.82 (dt, J=12.2, 7.6 Hz, 1H), 6.63-6.76(m, 2H), 6.84 (t, J=7.3 Hz, 1H), 7.14-7.38 (m, 5H), 7.44-7.69 (m, 3H),7.82-8.04 (m, 2H), 8.69 (d, J=8.3 Hz, 1H), 10.05 (s, 1H).

Example 5

Example 5 Step a

A mixture of ethyl 3-chloro-5-(trifluoromethyl)picolinate (1.336 g, 5.27mmol) and hydrazine hydrate (2 mL, 41.2 mmol) in ethanol (4 ml) washeated at 70° C. for 30 min (confirmed by TLC). After cooling, thereaction diluted with DCM (30 mL), washed with water and separated. Theaqueous layer was extracted with DCM. The combined organic layer waswashed with water and brine. The organic layer was dried (Na₂SO₄),filtered and concentrated to dryness. The residue was directly used fornext reaction. ESI-MS m/z=240.2 [M+H]⁺.

Example 5 Step b

A mixture of 3-chloro-5-(trifluoromethyl)picolinohydrazide (1.216 g,5.08 mmol) and morpholine (6 ml, 68.6 mmol) was heated at 110° C. for 4hrs. The reaction was concentrated, diluted with DCM (20 mL), washedwith water and separated. The aqueous layer was extractd with DCM. Thecombined organic layer was washed with H₂O and brine. The organic layerwas dried (Na₂SO₄), filtered and concentrated to dryness. The residuewas dissolved in EtOAc (8.4 mL, 7 vols) by instant heating. Then, theseed was added and kept at room temperature for 4 days (needle crystalformed). It was fitered through a fritted funnel, washed with 20% EtOAcin hexanes and dried. 1^(st) crystal: 630 mg. The mother liquor wasconcentrated to ˜⅓, kept for overnight, filtered, washed with 20% EtOAcin hexanes and dried to give 2^(nd) crystal: 162.6 mg. It was identicalto 1st one (by HPLC and proton-NMR). Total: 792.6 mg as a light yellowsolid. ESI-MS m/z=291.0 [M+H⁺]. ¹H-NMR (300 MHz, DMSO-d₆): δ 9.63 (s,1H), 8.49 (s, 1H), 7.71 (s, 1H), 4.54 (m, 2H), 3.77-3.68 (m, 4H),3.18-3.06 (m, 4H).

Example 5 Step c

To a oven-dried round-bottomed flask, CDI (960 mg, 5.92 mmol) wasdissolved in acetonitrile (9.87 mL) under nitrogen. The reaction mixturewas cooled to 0° C. and(S)-3-amino-5-phenyl-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-one(500 mg, 1.974 mmol) was added portionwise over 5 minutes. The reactionmixture was stirred at 0° C. for 1 hour and then removed from the icebath. The reaction mixture was warmed to RT and stirred for 2 hours.Next, water (213 μl, 11.84 mmol) was added dropwise with syringe. Thereaction mixture was concentrated to a viscous oil, which was takenforward crude. ESI-MS m/z=348.2 [M+H]⁺.

Example 5 Step d

In a oven-dried vial,(S)—N-(4-oxo-1-phenyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-3-yl)-1H-imidazole-1-carboxamide(274 mg, 0.789 mmol) was dissolved in DMSO (1578 μl). At roomtemperature, 3-morpholino-5-(trifluoromethyl)picolinohydrazide (229 mg,0.789 mmol) was added to the vial. The reaction mixture was allowed tostir at room temperature for 24 hours. Upon reaction completion, waterwas added to the flask and the aqueous was extracted with EtOAc (3×).The organic layer was dried with Na₂SO₄, filter and concentrated.(S)-2-(3-morpholino-5-(trifluoromethyl)picolinoyl)-N-(4-oxo-1-phenyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-3-yl)hydrazine-1-carboxamide(425 mg). ESI-MS m/z=570.2 [M+H]⁺.

Example 5 Step e

In a oven-dried vial,(S)-2-(3-morpholino-5-(trifluoromethyl)picolinoyl)-N-(4-oxo-1-phenyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-3-yl)hydrazine-1-carboxamide(449 mg, 0.788 mmol) was dissolved in dichloromethane (15.767 mL). Thereaction mixture was cooled to 0° C. and triethylamine (0.275 mL, 1.971mmol) and tosyl-Cl (225 mg, 1.183 mmol) were added, respectively. Thereaction mixture was warmed to room temperature and stirred overnight.The reaction was quench with NaHCO3 (sat.) and the aqueous layer wasextracted with DCM (3×). The organic layer was dried with Na₂SO₄,filtered and concentrated. The crude product was added to a silica gelcolumn and was eluted with acetone/hexane 0% to 100% to give(S)-3-((5-(3-morpholino-5-(trifluoromethyl)pyridin-2-yl)-1,3,4-oxadiazol-2-yl)amino)-5-phenyl-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-one(153 mg) as a yellow solid. The enantiopurity was enriched through HPLCchiral separation. (Column=CHIRALPAK IA-3, 5*0.46 cm (5 uM); MobilePhase=50% EtOH/50% hexanes (0.1% diethylamine); Flow rate=1 mL/min).ESI-MS m/z=552.2 [M+H]⁺.

Example 6

Example 6 Step a

A solution of ethyl 2-chloropyridine-3-carboxylate (1 g, 5.39 mmol, 1equiv.), morpholine (1.2 g, 13.8 mmol, 2.5 equiv.) in DMSO (5 mL) wasstirred at 100° C. for 1.5 hours. The mixture was cooled to r.t anddiluted with water, extracted with EA (3×). The combined organic phasewas washed with water and brine then dried, evaporated to give the titlecompound (1.48 g, crude). as yellow oil ESI-MS m/z=237.05 [M+H]⁺.

Example 6 Step b

A solution of ethyl 2-morpholinonicotinate (1.45 g, 6.14 mmol, 1equiv.), hydrazine hydrate (8 mL) in EtOH (8 mL) was stirred at 85° C.for 1 hour. The resulting mixture was concentrated under vacuum. Theresidue was purified by silica gel column chromatography, eluted withCH₂Cl₂/MeOH (0 to 10:1) to afford the title compound (1.28 g) as ayellow solid. ESI-MS m/z=223.00 [M+H]⁺.

Example 6 Step c

A solution of TCDI (213.8 mg, 1.2 mmol, 1.2 equiv.) and(S)-3-amino-5-phenyl-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-one(253 mg, 1.0 mmol, 1.0 equiv.) in DMF (2 mL) was stirred at r.t for 15minutes. Then 2-morpholinonicotinohydrazide (333 mg, 1.5 mmol, 1.5equiv.) was added. The mixture was stirred at r.t for 1 hour and dilutedwith water. The precipitated solids were collected by filtration andwashed with water to afford semicarbazide (336 mg, crude) as a yellowsolid. The crude semicarbazide (336 mg, 0.649 mmol, 1 equiv.) and EDCI(620 mg, 3.25 mmol, 5 equiv.) in DMF (4 mL) was stirred at 60° C. for1.5 hours. The resulting mixture was diluted with water. Theprecipitated solids were collected by filtration and washed with water.The crude product was purified by Prep Chiral-HPLC to afford the titlecompound (53.1 mg) as off white solid. ESI-MS m/z=484.30 [M+H]⁺. ¹H-NMR(300 MHz, DMSO-d₆): δ 3.14 (s, 4H), 3.68 (d, J=4.6 Hz, 4H), 3.98 (d,J=11.6 Hz, 1H), 4.21 (d, J=10.3 Hz, 1H), 6.71 (d, J=8.2 Hz, 2H), 6.87(d, J=7.3 Hz, 1H), 7.05 (, 4.8 Hz, 1H), 7.24 (m, 5H), 7.96 (m, 1.8 Hz,1H), 8.31 (d, J=9.1 Hz, 1H), 8.36 (m, 1H), 10.13 (s, 1H).

Example 7

Example 7 Step a

A solution of 5-bromo-3-fluoropyridine-2-carboxylic acid (2.7 g, 12.27mmol, 1 equiv.) and H₂SO₄ (5 mL) in MeOH (10 mL) was stirred at 85° C.for 1 hour. The mixture was basified to pH 7 with saturated NaHCO₃. Theresulting mixture was extracted with EA (3×) and evaporated to affordthe title compound (2.3 g) as off white solid. ESI-MS m/z=233.90 [M+H]⁺.

Example 7 Step b

A solution of methyl 5-bromo-3-fluoropicolinate (1.7 g, 19.79 mmol, 2equiv.), K₃PO₄ (4.6 g, 19.79 mmol, 2 equiv), Pd(dppf)Cl₂ (230 mg) andH₂O (0.5 mL) in toluene (23 mL). The resulting solution was stirred at100° C. for 1 hour. The mixture was cooled to room temperature andextracted with EA (3×). The combined organic phase was evaporated toafford the title compound (1.48 g) as yellow oil. ESI-MS m/z=195.95[M+H]⁺.

Example 7 Step c

A solution of methyl 5-cyclopropyl-3-fluoropicolinate (1.3 g, 6.66 mmol,1 equiv.), morpholine (1.2 g, 13.33 mmol, 2 equiv.) and DIPEA (1.7 g,13.33 mmol, 2 equiv.) in DMSO (8 mL) was stirred at 120° C. for 2 hours.The resulting mixture was extracted with EA (3×) and washed with water,dried over Na₂SO₄. After filtration, the filtrate was concentrated underreduced pressure. The residue was purified by silica gel columnchromatography, eluted with EA/PE (5:1) to afford methyl5-cyclopropyl-3-morpholinopicolinate as yellow solid (1.2 g). ESI-MSm/z=263.00 [M+H]⁺.

Example 7 Step d

A solution of methyl 5-cyclopropyl-3-morpholinopicolinate (500 mg, 1.91mmol, 1 equiv.) and hydrazine hydrate (4 mL) in EtOH (4 mL) was stirredat 85° C. for 2 hours. The resulting mixture was concentrated undervacuum. The residue was purified by silica gel column chromatography,eluted with CH₂Cl₂/MeOH (0 to 10:1) to afford product (450 mg) as brownoil. ESI-MS m/z=263.05 [M+H]⁺.

Example 7 Step e

Prepared in a similar fashion to example 6 step c using5-cyclopropyl-3-morpholinopicolinohydrazide. ESI-MS m/z=524.25 [M+H]⁺.¹H-NMR (300 MHz, DMSO-d₆): δ 0.82-0.94 (m, 2H), 1.01-1.14 (m, 2H),1.95-2.12 (m, 1H), 2.96 (m, 4H), 3.70 (t, J=4.5 Hz, 4H), 3.87-4.01 (m,1H), 4.19 (m, 1H), 4.46 (m, 1H), 6.70 (d, J=8.1 Hz, 2H), 6.85 (m, 1H),7.12-7.38 (m, 8H), 8.13 (d, J=1.8 Hz, 1H), 8.30 (d, J=9.3 Hz, 1H), 10.11(s, 1H).

Example 8

Example 8 Step a

A solution of ethyl 3-chloro-5-(trifluoromethyl)pyridine-2-carboxylate(3 g, 11.86 mmol, 1 equiv.), DIPEA (3.1 g, 23.72 mmol, 2 equiv.) andmethoxyethan-1-amine (0.9 g, 11.86 mmol, 1 equiv.) in DMSO was stirredat 100° C. overnight. The mixture was cooled to room temperature anddiluted with water, extracted with EA (3×). The combined organic phasewas washed with water and brine then dried, evaporated. The residue waspurified by silica gel column chromatography, eluted with EA/PE (0 to1:1) to afford ethyl3-((2-methoxyethyl)amino)-5-(trifluoromethyl)picolinate (700 mg) asyellow solid. ESI-MS m/z=293.10 [M+H]⁺.

Example 8 Step b

A solution of ethyl3-((2-methoxyethyl)amino)-5-(trifluoromethyl)picolinate (500 mg, 2 mmol,1 equiv.) and hydrazine hydrate (8 mL) in EtOH (8 mL) was stirred at 85°C. for 1.5 hours. The resulting mixture was concentrated under vacuum.The residue was purified by silica gel column chromatography, elutedwith CH₂Cl₂/MeOH (0 to 10:1) to afford product (480 mg) as yellow solid.ESI-MS m/z=278.95 [M+H]⁺.

Example 8 Step c

Prepared in a similar fashion to Example 6, step c using3-((2-methoxyethyl)amino)-5-(trifluoromethyl)picolinohydrazide. ESI-MSm/z=540.15 [M+H]⁺. ¹H-NMR (300 MHz, DMSO-d₆) δ 3.54 (m, 2H), 3.59 (d,J=4.7 Hz, 2H), 3.88-4.02 (m, 1H), 4.23 (m, 6.6 Hz, 1H), 4.43-4.58 (m,1H), 6.71 (d, J=8.2 Hz, 2H), 6.86 (m, 1H), 7.12-7.37 (m, 6H), 7.59 (d,J=1.8 Hz, 1H), 7.79 (m, 1H), 8.25 (d, J=1.7 Hz, 1H), 8.68 (d, J=9.1 Hz,1H), 10.15 (s, 1H).

Example 9

Example 9 Step a

The solution of 5-methylfuran-2-carboxylic acid (1 g, 7.93 mmol, 1equive.), H₂SO₄ (2 mL) in MeOH (10 mL) was stirred at 85° C. for 1 hour.The pH value of the solution was adjusted to 7 with Na₂CO₃. Theresulting solution was extracted with EA (3×) and evaporated to affordproduct (900 mg, crude) as brown solid and used directly for next step.

Example 9 Step b

A solution of methyl 5-methylfuran-2-carboxylate (900 mg, crude, stepa), hydrazine hydrate (4 mL) in EtOH (4 mL) was stirred at 85° C. for 1hour. The resulting mixture was concentrated under vacuum. The residuewas purified by silica gel column chromatography, eluted withCH₂Cl₂/MeOH (0 to 10:1) to afford product (1.4 g, crude) as yellow oil.ESI-MS m/z=182.00 [M+ACN+H]⁺.

Example 9 Step c

Prepared in a similar fashion to Example 6, step c using5-methylfuran-2-carbohydrazide. ESI-MS m/z=402.25 [M+H]⁺. ¹H-NMR (300MHz, DMSO-d₆): δ 2.38 (s, 3H), 3.85-3.99 (m, 1H), 4.12-4.25 (m, 1H),4.40 (d, J=13.6 Hz, 1H), 6.35 (m, 1H), 6.69 (d, J=8.2 Hz, 2H), 6.85 (t,J=7.4 Hz, 1H), 6.95 (d, J=3.5 Hz, 1H), 7.25 (m, 6H), 8.33 (d, J=9.2 Hz,1H), 10.12 (s, 1H).

Example 10

Example 10 Step a

The solution of ethyl1-cyclopentyl-3-morpholino-1H-pyrazole-4-carboxylate (300 mg, 1.02 mmol,1 equiv.), hydrazine hydrate (3 mL) in EtOH (3 mL) was stirred at 85° C.for 12 hours. The resulting mixture was concentrated under vacuum. Theresidue was purified by silica gel column chromatography, eluted withCH₂Cl₂/MeOH (0 to 10:1) to afford1-cyclopentyl-3-morpholino-1H-pyrazole-4-carbohydrazide (360 mg, crude)as yellow solid. ESI-MS m/z=280.15 [M+H]⁺.

Example 10 Step b

Prepared in a similar fashion to Example 6, step c using1-cyclopentyl-3-morpholino-1H-pyrazole-4-carbohydrazide. ESI-MSm/z=540.15 [M+H]⁺. ¹H-NMR (300 MHz, DMSO-d₆): δ 1.64 (m, 2H), 1.73-1.84(m, 2H), 1.92 (m, 2H), 2.03 (m, 2H), 3.16 (s, 4H), 3.68 (m, 4H),3.87-4.01 (m, 1H), 4.16 (m, 1H), 4.39 (m, 1H), 4.63 (m, 1H), 6.69 (d,J=8.1 Hz, 2H), 6.85 (m, 1H), 7.11-7.37 (m, 6H), 8.03 (d, J=9.3 Hz, 1H),8.05 (s, 1H), 10.09 (s, 1H).

Example 11

Example 11 Step a

A solution of methyl 4-fluorobenzoate (3.4 g, 22.06 mmol, 1 equiv.) andhydrazine hydrate (10 mL) in EtOH (10 mL) was stirred at 85° C. for 1hour. The resulting mixture was concentrated under vacuum. The residuewas purified by silica gel column chromatography, eluted withCH₂Cl₂/MeOH (0 to 10:1) to afford product (2.5 g) as a white solid.ESI-MS m/z=540.15 [M+H]⁺. ESI MS m/z=154.95 [M+H]⁺.

Example 11 Step b

Prepared in a similar fashion to Example 6, step c using4-fluorobenzohydrazide. ESI-MS m/z=416.20 [M+H]⁺. ¹H-NMR (300 MHz,DMSO-d₆): δ 3.90-4.00 (m, 1H), 4.20 (m, 1H), 4.40-4.52 (m, 1H), 6.70 (d,J=8.0 Hz, 2H), 6.85 (m, 1H), 7.13-7.34 (m, 6H), 7.41 (m, 2H), 7.81-7.93(m, 2H), 8.35 (d, J=9.1 Hz, 1H), 10.13 (s, 1H).

Example 12

Example 12 Step a

A solution of methyl 4-bromo-2-fluorobenzoate (1 g, 4.29 mmol, 1equiv.), morpholine (748 mg, 8.58 mmol, 2 equiv.), Ruphos (300 mg),Cs₂CO₃ (1.6 g, 5.06 mmol, 1.2 equiv.) and Ruphos-2nd-precatlyst (130 mg)in 1,4-dioxane (10 mL) was stirred at 100° C. for 1 hour. The resultingmixture was extracted with EA (3×) and combined organic phase, then itwas concentrated under vacuum to give the residue. The residue waspurified by silica gel column chromatography, eluted with EA/PE (1:3) toafford methyl 2-fluoro-4-morpholinobenzoate (840 mg) as a yellow solid.ESI-MS m/z=240.10 [M+H]⁺.

Example 12 Step b

A solution of methyl 2-fluoro-4-morpholinobenzoate (840 mg, 3.51 mmol, 1equiv.) and hydrazine hydrate (2 mL) in EtOH (2 mL) was stirred at 85°C. overnight. The resulting mixture was concentrated under vacuum. Theresidue was purified by silica gel column chromatography, eluted withCH₂Cl₂/MeOH (0 to 10:1) to afford the title compound (540 mg) as a darkgray solid. ESI-MS m/z=240.50 [M+H]⁺.

Example 12 Step c

Prepared in a similar fashion to Example 6, step c using2-fluoro-4-morpholinobenzohydrazide. ESI-MS m/z=501.25 [M+H]⁺. ¹H-NMR(300 MHz, DMSO-d₆): δ 3.28 (d, J=9.9 Hz, 4H), 3.74 (m, 4H), 3.94 (m,1H), 4.18 (m, 1H), 4.35-4.51 (m, 1H), 6.65-6.75 (m, 2H), 6.79-6.98 (m,3H), 7.23 (m, 6H), 7.64 (m, 1H), 8.23 (d, J=9.2 Hz, 1H), 10.11 (s, 1H).

Example 13

Example 13 Step a

A solution of (E)-ethyl 2-cyano-3-ethoxyacrylate (1.69 g, 0.01 mol) inTHF (5 mL) was dropwised to the solution of 2-hydrazinylpyridine (1.439g, 0.013 mol) and NaOEt-EtOH (8.56 g, 0.026 mol) at 0° C. The mixturewas stirred for 1.5 hours at 0° C. before quenched with 4 M HCl in1,4-dioxane (6.5 mL, 0.026 mol). The solution was refluxed for 2 hours.It was adjusted pH=10-13 with 1 M NaOH and concentrated. The crudeproduct was purified by Flash-Prep-HPLC (MeCN/H₂O) to give desiredcompound as an orange solid (1.07 g). ESI-MS m/z=233.25 [M+H]⁺. ¹H-NMR(300 MHz, DMSO-d₆): δ 1.29 (m, 3H), 4.24 (m, 2H), 7.32 (m, 1H), 7.71 (m,1H), 7.97 (m, 1H), 8.43 (m, 1H), 8.67 (s, 1H).

Example 13 Step b

NaH (202 mg, 8.4 mmol) was added to the solution of ethyl3-amino-1-(pyridin-2-yl)-1H-pyrazole-4-carboxylate (970 mg, 4.2 mmol) inDMF (5 mL) at 0° C. The mixture was stirred for 10 minutes at 0° C.1-bromo-2-(2-bromoethoxy)ethane (1.923 g, 8.4 mmol) was added, and thesolution was stirred overnight. The mixture was quenched with water,extracted with EA (3×). The organic layers were combined and washed withbrine (2×), dried and concentrated. The residue was chromatographied(silica, petroleum ether-ethyl acetate) to give desired compound as redoil (340 mg). ESI-MS m/z=303.30 [M+H]⁺.

Example 13 Step C

A solution of ethyl3-morpholino-1-(pyridin-2-yl)-1H-pyrazole-4-carboxylate (340 mg, 1.122mmol) and NH₂NH₂.H₂O (3 mL) in EtOH (4 mL) was refluxed overnight. Thecrude product was purified by Flash (MeCN/H₂O) to give desired compoundas a white solid (280 mg). ESI-MS m/z=311.25 [M+H]⁺.

Example 13 Step d

Prepared in a similar fashion to Example 5 step d using3-morpholino-1-(pyridin-2-yl)-1H-pyrazole-4-carbohydrazide. ESI-MSm/z=568.2 [M+H]⁺.

Example 13 Step e

Prepared in a similar fashion to Example 5 step d using(S)-2-(3-morpholino-1-(pyridin-2-yl)-1H-pyrazole-4-carbonyl)-N-(4-oxo-1-phenyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-3-yl)hydrazine-1-carboxamide.ESI-MS m/z=550.3 [M+H]⁺.

Example 14

Example 14 Step a

In an oven-dried vial, ethyl 5-bromothiazole-4-carboxylate (200 mg, 0.88mmol) was dissolved in MeCN (2.4 mL). Morpholine (87 uL, 0.99 mmol) andDBU (0.2 mL, 1.35 mmol) were added to the vial sequentially. The vialwas sealed and heated to 80° C. for 5 hours. Cool the vial to roomtemperature and quench with water. Extract aqueous layer (3×) withEtOAc. The organic layer was dried with NaSO₄, filtered andconcentrated. The crude product was purified on silica gel(hexane/EtOAc: 0% to 80%), affording ethyl5-morpholinothiazole-4-carboxylate (120 mg) as a white solid. ESI-MSm/z=243.1 [M+H]⁺.

Example 14 Step b

To an oven-dried vial, ethyl 5-morpholinothiazole-4-carboxylate (247 mg,1.06 mmol) was dissolved in MeCN (5.4 mL). N-bromosuccimide (208 mg,1.17 mmol) was added to the vial in one portion at room temperature. Thereaction was allowed to stir at room temperature until the startingmaterial was consumed. The reaction mixture was concentrated andpurified on silica gel (hexane/EtOAc: 0% to 80%), affording ethyl2-bromo-5-morpholinothiazole-4-carboxylate (256 mg) as a white solid.ESI-MS m/z=322.2 [M+H]⁺.

Example 14 Step c

To an oven-dried vial, ethyl 2-bromo-5-morpholinothiazole-4-carboxylate(650 mg, 2.02 mmol) was dissoved in DMF (10 mL).(1,10-Phenanthroline)(trifluoromethyl)copper (760 mg, 2.43 mmol) wasadded in one portion and the reaction mixture was heated at 80° C.overnight. The reaction mixture was cooled to room temperature anddiluted with EtOAc. The organic layer was washed with H₂O (3×), brineand then dried with Na₂SO₄. The solid was filtered and the organic layerwas concentrated to dryness. The residue was purified by columnchromatography with 0-20% EtOAc in hexanes to give ethyl5-morpholino-2-(trifluoromethyl)thiazole-4-carboxylate (360 mg). ESI-MSm/z=311.0 [M+H]⁺.

Example 14 Step d

To a round-bottomed flask, ethyl5-morpholino-2-(trifluoromethyl)thiazole-4-carboxylate (432 mg, 1.39mmol) was dissolved in ethanol (4.64 mL) and hydrazine hydrate (2.32 mL)open to air to give a red solution. The reaction mixture was heated to70° C. for 1 hour then cooled to room temperature. The reaction mixturewas concentrated in vacuo. The crude product was added to a silica gelcolumn and was eluted with methanol/dichloromethane 0% to 10% to give5-morpholino-2-(trifluoromethyl)thiazole-4-carbohydrazide (367 mg) as asolid. ESI-MS m/z=297.1 [M+H]⁺.

Example 14 Step e

Prepared in a similar fashion to Example 6 step d using5-morpholino-2-(trifluoromethyl)-thiazole-4-carbohydrazide. ESI-MSm/z=576.2 [M+H]⁺.

Example 14 Step f

To a mixture of(S)-2-(5-morpholino-2-(trifluoromethyl)thiazole-4-carbonyl)-N-(4-oxo-1-phenyl-2,3,4,5-tetrahydro-1H-benzo[b][1,4]diazepin-3-yl)hydrazine-1-carboxamide(72 mg, 0.13 mmol) and triethylamine (52.3 μl, 0.38 mmol) in DCM (1.2mL) was added phosphoryl trichloride (23.3 μl, 0.25 mmol) at 0° C. andstirred at room temperature for 15.5 hrs. The reaction was diluted withEtOAc, and then washed with H₂O, sat. NaHCO₃ solution and brine. Theorganic layer was dried (Na₂SO₄), filtered and concentrated to dryness.The residue was purified by silica gel column with 0˜100% acetone inhexane to give(S)-3-((5-(5-morpholino-2-(trifluoromethyl)thiazol-4-yl)-1,3,4-oxadiazol-2-yl)amino)-5-phenyl-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-one(2.6 mg). ESI-MS m/z=558.2 [M+H]⁺.

Example 15

Example 15 Step a

To a vial, add ethyl 2-bromo-5-morpholinothiazole-4-carboxylate (212 mg,0.67 mmol), cyclopropylboronic acid (65 mg, 0.76 mmol), K₂CO₃ (286 mg,2.07 mmol) and Pd(PPh₃)₄ (40 mg, 0.035 mmol). The vial was sealed andevacuated with nitrogen. Toluene (2.88 mL) and water (0.58 mL) wereadded to the vial with a syringe. The reaction mixture was heated to 80°C. and stirred at that temperature for 20 hours. The vial was cooled toroom temperature and quenched with water. The aqueous layer wasextracted (3×) with EtOAc. The organic layer was dried with Na₂SO₄,filtered and concentrated. The crude product was added to a silica gelcolumn and was eluted with ethyl acetate/hexane 0% to 100% to give ethyl2-cyclopropyl-5-morpholinothiazole-4-carboxylate (76 mg) as a solid.ESI-MS m/z=283.1 [M+H]⁺.

Example 15 Step b

Prepared in a similar fashion to Example 15, step d using ethyl2-cyclopropyl-5-morpholinothiazole-4-carboxylate. ESI-MS m/z=269.2[M+H]⁺.

Example 15 Step c

To a solution of(S)-3-amino-5-phenyl-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-one(47.2 mg, 0.186 mmol) in DMF (1165 μl) was added1,1′-thiocarbonyldiimidazole (39.8 mg) and stirred for 1 hour. After anhour, 2-cyclopropyl-5-morpholinothiazole-4-carbohydrazide (50 mg, 0.186mmol) and DMF (1165 μl) were added and the mixture was stirred for 2hours. EDC (179 mg, 0.932 mmol) was added and the mixture was stirredfor 4 hours at 60° C. The reaction mixture was diluted with water andextracted with EtOAc (3×). The organic layer was dried, filtered andconcentrated. The crude product was added to a silica gel column and waseluted with acetone/hexane 0% to 50% to give the racemate as a yellowsolid. The racemic mixture was purified by chiral separation.(Column=YMC CHIRAL Cellulose-SB, 250*20 mm (5 uM); Mobile Phase=50%iPrOH/50% hexanes; Flow rate=20 mL/min). ESI-MS m/z=530.2 [M+H]⁺.

Example 16

Example 16 Step a

Ethyl 2-bromo-5-morpholinothiazole-4-carboxylate (1 g, 3.11 mmol),pyridin-3-yl boronic acid (0.574 g, 4.67 mmol), Cs₂CO₃ (2.029 g, 6.23mmol), and Pd(DPPF)Cl₂ (0.254 g, 0.311 mmol) were added to a oven-driedvial and sealed. The vial was evacuated and refilled with nitrogen. DMF(12.45 ml) was added through the septum via a syringe and the vial washeated to 90° C. overnight. The reaction mixture was cooled to roomtemperature and filtered through celite. The filtrate was concentratedin vacuo and the resultant crude residue was purified using columnchromatography on silica gel (eluent: Hexane/EtOAc (4:1)) to provideethyl 5-morpholino-2-(pyridin-3-yl)thiazole-4-carboxylate (222 mg).ESI-MS m/z=320.1 [M+H]⁺.

Example 16 Step b

Prepared in a similar fashion to Example 15, step d using ethyl5-morpholino-2-(pyridin-3-yl)thiazole-4-carboxylate. ESI MS m/z=306.2[M+H]⁺.

Example 16 Step c

Prepared in a similar fashion to Example 15 step c using ethyl5-morpholino-2-(pyridin-3-yl)thiazole-4-carboxylate. ESI-MS m/z=567.2[M+H]⁺.

Example 17

Example 17 Step a

Prepared in a similar fashion to Example 1 using5-bromo-1,2,3-trifluorobenzene. ESI-MS m/z=308.2 [M+H]⁺.

Example 17 Step b

Prepared in a similar fashion to Example 15 step c using(S)-3-amino-5-(3,4,5-trifluorophenyl)-1,3,4,5-tetrahydro-2H-benzo[b][1,4]diazepin-2-oneand 3-morpholino-5-(trifluoromethyl)picolinohydrazide. ESI-MS m/z=606.3[M+H]⁺.

Example 18

Example 18 Step a

A solution of 3-bromoquinoline (2.0 g, 9.66 mmol, 1.0 equiv.), Ruphos(600 mg, 1.28 mmol, 0.13 equiv.), NaOBu-t (1.1 g, 11.6 mmol, 1.2 equiv.)and Ruphos 2^(nd) gen Pd precatlyst (372.8 mg, 0.48 mmol, 0.05 equiv.)in morpholine (30 mL) was stirred at 110° C. for 5 hours and then cooledto room temperature. The resulting mixture was extracted with EtOAc (3×)and the combined organic phase was dried over anhydrous Na₂SO₄,concentrated under vacuum. The residue was purified by silica gel columnchromatography, eluted with EtOAc/PE (1:3) to afford4-(quinolin-3-yl)morpholine (1.0 g, 4.9 mmol, 50%) as a yellow solid.ESI-MS m/z=215.20 [M+H]⁺.

Example 18 Step b

A solution of 4-(quinolin-3-yl)morpholine (1.0 g, 4.67 mmol, 1.0 equiv.)in DCM (30 mL) was added AlCl₃ (13.1 mg, 0.098 mmol, 0.013 equiv.).Benzoyl chloride (1.06 g, 7.56 mmol, 1.62 equiv.) was added dropwise.The mixture was stirred for 2 hours at room temperature and TMSCN (1.0g, 7.5 mmol, 1.62 equiv.) was added. The resulting mixture was stirredovernight at room temperature. The mixture was diluted with DCM, washedwith 1N HCl aqueous solution, saturated Na₂CO₃ aqueous solution, driedover anhydrous Na₂SO₄ and concentrated. The residue was purified bysilica gel column chromatography, eluted with PE/EtOAc (0 to 5:1) toafford 1-benzoyl-3-morpholino-1,2-dihydroquinoline-2-carbonitrile (400mg, 1.16 mmol, 25%) as a yellow solid.

Example 18 Step c

A solution of 1-benzoyl-3-morpholino-1,2-dihydroquinoline-2-carbonitrile(400 mg, 1.16 mmol, 1 equiv.) in formic acid (10 mL) was stirred for 2hours at room temperature. The resulting mixture was adjusted pH to 6with saturated NaHCO₃ aqueous solution and extracted with EtOAc (3×).The combined organic phase was evaporated to afford3-morpholinoquinoline-2-carbonitrile (250 mg, 1.05 mmol, 90%) as brownoil. ESI-MS m/z=240.10 [M+H]⁺.

Example 18 Step d

A solution of 3-morpholinoquinoline-2-carbonitrile (250 mg, 1.05 mmol, 1equiv.), KOH (291 mg, 2.1 mmol, 2 equiv.) and H₂O (2 mL) in EtOH (3 mL)was stirred for 6 hours at 100° C. The resulting mixture was cooled toroom temperature, adjusted pH to 4 with HCl and extracted with EtOAc(3×). The combined organic phase was evaporated and purified by reversephase column to afford 3-morpholinoquinoline-2-carboxylic acid (70 mg,0.27 mmol, 26%) as yellow solid. ESI-MS m/z=259.25 [M+H]⁺.

Example 18 Step e

A solution of 3-morpholinoquinoline-2-carboxylic acid (70 mg, 0.27 mmol,1 equiv.), tert-butyl hydrazinecarboxylate (36 mg, 0.27 mmol, 1 equiv.),DIPEA (35 mg, 0.54 mmol, 2 equiv.) and HATU (123 mg, 0.32 mmol, 1.2equiv.) in DMF (3 mL) was stirred for 1 hour at room temperature. Theresulting mixture was extracted with EtOAc (3×) and washed by water(3×). Then the combined organic phase was evaporated and purified byPrep-TLC to afford tert-butyl2-(3-morpholinoquinoline-2-carbonyl)hydrazine-1-carboxylate (148 mg,crude) as brown oil. ESI-MS m/z=373.05 [M+H]⁺.

Example 18 Step f

A solution of tert-butyl2-(3-morpholinoquinoline-2-carbonyl)hydrazine-1-carboxylate (148 mg,0.40 mmol, 1 equiv.) and HCl (0.25 mL) in EtOAc (2 mL) was stirred atroom temperature for 1 hour. The resulting mixture was adjusted pH to 7with saturated NaHCO₃ aqueous solution, extracted with DCM (3×), driedover anhydrous Na₂SO₄. The combined organic phase was evaporated toafford 3-morpholinoquinoline-2-carbohydrazide (120 mg, crude) as brownoil. ESI-MS m/z=273.05 [M+H]⁺.

Example 18 Step g

Prepared in a similar fashion to Example 15 step c using3-morpholinoquinoline-2-carbohydrazide. ESI-MS m/z=534.30 [M+H]⁺. ¹H NMR(300 MHz, DMSO-d6) δ 3.05 (s, 4H), 3.77 (s, 4H), 3.93-4.01 (m, 1H),4.18-4.27 (m, 1H), 4.52 (d, J=10.7 Hz, 1H), 6.72 (d, J=8.1 Hz, 2H), 6.86(t, J=7.3 Hz, 1H), 7.16-7.35 (m, 6H), 7.62-7.75 (m, 2H), 7.99 (d, J=8.1Hz, 2H), 8.10 (s, 1H), 8.56 (d, J=9.2 Hz, 1H), 10.16 (s, 1H).

Example 19

Example 19 Step a

To a mixture of the N¹-phenylbenzene-1,2-diamine (10.0 g, 54 mmol),DIPEA (14.0 g108 mmol) and 3-ethoxy-3-oxopropanoic acid (7.2 g, 54 mmol)in DMF (100 mL) was added HATU (41.8 g, 108 mmol). The mixture wasstirred at room temperature overnight. Then it was diluted with water,extracted by EtOAc and purified by silica gel column to afford the titlecompound as a red solid (8.0 g). ESI-MS m/z=299.25 [M+H]⁺.

Example 19 Step b

A solution of the compound from step a (7.0 g, 23.5 mmol) in THF/H₂O (40mL/40 mL) was added LiOH (2.3 g, 94.0 mmol). The reaction mixture wasstirred at RT for 1 hr. It was acidified by HCl, extracted by EA (×2),washed with saturated NaCl aqueous solution. The organic layer was driedand concentrated to give the desired compound as a white solid (6.2 g).ESI-MS m/z=271.20 [M+H]⁺.

Example 19 Step c

A solution of the compound from step b (6.2 g, 23 mmol), EDCI (8.8 g, 46mmol), HOBt (6.2 g, 46 mmol) in DMF (60 mL) was added DIEPA (5.9 g, 46mmol). Then the reaction mixture was stirred overnight at RT. It wasdiluted with EA, washed with water (×2). The organic layer was dried andpurified by recrystallization (PE/EA=1:1) to give the desired compoundas a white solid (2.2 g). ESI-MS m/z=253.20 [M+H]⁺.

Example 19 Step d

A solution of the compound from step c (2.2 g, 8.7 mmol), K₂CO₃ (2.4 g,17.4 mmol) in DMF (20 mL) was added PMBCl (1.5 g, 10.5 mmol). Then thereaction mixture was stirred at RT overnight. LCMS showed that thereaction was complete. It was diluted with EA, washed with water (×2).The organic layer was dried and purified by recrystallization(PE/EtOAc=5:1) to give the desired compound as a white solid (2.5 g).ESI-MS m/z=373.30 [M+H]⁺.

Example 19 Step e

A solution of the compound from step d (2.5 g, 6.7 mmol) in THF (100 mL)was added tert-BuOK (1.5 g, 13.4 mmol) at −78° C., the reaction mixturewas stirred for 30 min at −78° C. Then 2,4,6-triisopropylbenzenesulfonylazide (3.1 g, 10.1 mmol) in THF (50 mL) was added dropwise into thereaction mixture at −78° C. After the addition, the reaction mixture wasstirred at −78° C. for 10 mins. AcOH (1 mL) added and the reaction waswarmed to RT overnight. It was purified by silica gel column andconcentrated to give the title compound as a white solid (0.6 g). ESI-MSm/z=414.30 [M+H]⁺.

Example 19 Step f

A solution of the compound from step e (0.6 g, 1.5 mmol) in MeOH/DCM (30mL/30 mL) was added Pd/C (20 mg) at RT. Then the reaction was replacedH₂ three times and stirred for 30 min. The reaction mixture was filteredand the cake was washed with MeOH (×3), the filtrate was concentrated togive the title compound as a white solid (500 mg). ESI-MS m/z=388.35[M+H]⁺.

Example 19 Step g

A solution of the compound from step f (500 mg, 1.3 mmol) in MeCN/H₂O (8mL/2 mL) was added CAN (7.1 g, 13 mmol) at 0° C. The reaction mixturewas stirred at RT for 1 hr. It was diluted with H₂O, washed with EtOAc(×3), the aqueous phase was adjust pH=9 with saturated aqueous NaHCO₃solution, then extracted with EA (×3) and the combined layer wasconcentrated to give the title compound as a yellow solid (180 mg).ESI-MS m/z=268.15 [M+H]⁺.

Example 19 Step h

A solution of the compound from step g (60 mg, 0.23 mmol),1,1′-thiocarbonyldiimidazole (48 mg, 0.27 mmol) in DMF (5 mL) wasstirred for 30 minutes before benzohydrazide (100 mg, 0.69 mmol) wasadded. And it was stirred for 30 minutes. It was concentrated to givethe title compound as yellow oil, which was used directly for the nextstep. ESI-MS m/z=446.05 [M+H]⁺.

Example 19 Step i

A solution of the compound from step h (crude) and EDCI (52 mg, 0.27mmol) in DMF (5 mL) was stirred for 30 minutes at 60° C. It was purifiedby Prep-HPLC (MeCN/H₂O) to give the title compound as a light solid(20.3 mg). ESI-MS m/z=412.10 [M+H]⁺. ¹H NMR (300 MHz, DMSO-d₆): δ 5.09(d, J=7.8 Hz, 1H), 6.97 (d, J=8.2 Hz, 1H), 7.17-7.29 (m, 3H), 7.32-7.44(m, 3H), 7.49 (m, 2H), 7.53-7.63 (m, 3H), 7.80-7.89 (m, 2H), 8.58 (d,J=8.2 Hz, 1H), 11.09 (s, 1H).

Example 20

Example 20 was prepared from the compound (step g in example 18) andphenyl isocyanate. ESI-MS m/z=387.30 [M+H]⁺.

Example 21

A solution of the compound from step gin example 19 (60 mg, 0.23 mmol),TEA (47 mg, 0.43 mmol) in DCM (2 mL) was added BzCl (39 mg, 0.28 mmol)at RT. The reaction mixture was stirred at RT for 1 hr. LCMS showed thatthe starting material was consumed. It was purification by prep-HPLC togive desired compound as a white solid (18.5 mg). ESI-MS m/z=372.30[M+H]⁺.

Assays

Introduction

RSV is a single stranded negative sense RNA virus that causesrespiratory tract infections which can be dangerous to infants, theelderly, and immunosuppressed individuals. Currently there is novaccine, and therapeutic options are both costly and of limitedeffectiveness. These approved treatments are Ribavirin, andPalivizumab/Synagis (a monoclonal antibody). RSV has two genotypes, Aand B, which differ primarily in the structure of the virus' surface “G”attachment protein. Our current primary screen focusses on RSV-A anduses an in vitro cytoprotection assay where compounds are added in2-fold dilutions to cells which are then subjected to fully replicativeviral particles. Cell viability is measured several days later alongwith separate measurements of compound cytotoxicity. This report focuseson the results of our most recent screening of compounds.

Methods

HEp-2 cells, (originally derived from tumors grown inirradiated-cortisonised weanling rats that had been injected withepidermoid carcinoma tissue from a 56 year old male's larynx, but laterfound to be indistinguishable from HeLa cells by PCR DNA analysis), wereused for the culturing of genotype A, “Long” strain RSV. Flasks wereinoculated with RSV and viral stocks were collected once cytopathiceffect (CPE) was greater than 90%. Viral stocks in 25% sucrose mediawere snap frozen using liquid nitrogen to increase viral stability.Viral stock titers were quantified by tissue culture infectious dose 50%(TCID50) using 8,000 cells per well and 3-fold viral dilutions across a96-well plate, cultured for 4 days.

Following extensive parameter testing, the final assay is run asfollows: HEp-2 cells are seeded into the inner 60 wells of a 96-wellplate at 8,000 cells per well in a volume of 50 using Growth Media (DMEMwithout phenol red, 1% L-Glut, 1% Penn/Strep, 1% nonessential aminoacids, 10% FBS). 2-fold serial dilutions of control and test compoundsare added to the wells in duplicate in a total volume of 25 μL. Viralstock is then added to the wells in a volume of 25 μL, bringing thetotal volume of each well to 100 μL. Each 96-well plate has a controlcolumn of 6 wells with cells and virus but no compound (negativecontrol, max CPE), a column with cells but no compound or virus(positive control, minimum CPE), and a column with no cells or virus orcompound (background plate/reagent control). The control wells withcells but no virus are given an additional 25 uL of growth mediacontaining an equal quantity of sucrose as those wells receiving theviral stock in order to keep consistent in media and volume conditions.The outer wells of the plate are filled with 125 μL of growth media toact as a thermal and evaporative moat around the test wells. Following a4-day incubation period, the plates are read using ATPlite (50 uL addedper well), which quantifies the amount of ATP (a measure of cell health)present in each well. Assay plates are read using the Envisionluminometer. These data are used to calculate the EC₅₀ of each compound.EC₅₀ ranges are as follows: A<2 μM; B 2-5 μM; C>5 μM.

TABLE 2 Summary of Activities Human RSV-A (“Long” strain) Example EC₅₀ 1C 2 C 3 C 4 C 5 A 6 B 7 A 8 C 9 C 10 A 11 C 12 C 13 B 14 A 15 A 16 A 17C 19 C 20 C 21 C

While this invention has been particularly shown and described withreferences to preferred embodiments thereof, it will be understood bythose skilled in the art that various changes in form and details may bemade therein without departing from the scope of the inventionencompassed by the appended claims.

What is claimed:
 1. A compound represented by Formula (I), or apharmaceutically acceptable salt thereof:

wherein: A is selected from the group consisting of: 1) Optionallysubstituted heteroaryl; and 2) Optionally substituted aryl; B isselected from the group consisting of: 1) Optionally substitutedheteroaryl; and 2) Optionally substituted aryl; L is —NH—; X is —CH₂—,or —C(O)—; R₁ is selected from the group consisting of: 1) Optionallysubstituted heteroaryl; and 2) Optionally substituted aryl; R₂ is absentor is selected from the group consisting of: 1) Optionally substituted—C₁-C₈ alkyl; 2) Optionally substituted —C₂-C₈ alkenyl; 3) Optionallysubstituted —C₂-C₈ alkynyl; 4) Optionally substituted —C₁-C₈ alkoxy; 5)Optionally substituted aryloxy; 6) Optionally substituted —C₃-C₁₂cycloalkyl; 7) Optionally substituted —C₃-C₁₂ cycloalkenyl; 8)Optionally substituted 3- to 12-membered heterocycloalkyl; 9) Optionallysubstituted aryl; 10) Optionally substituted arylalkyl; 11) Optionallysubstituted heteroaryl; 12) Optionally substituted heteroarylalkyl; 13)—NR₁₃R₁₄; 14) —CO—NR₁₃R₁₄; and 15) —SO₂—NR₁₃R₁₄; Each R₃ isindependently selected from halogen, hydroxyl, protected hydroxyl,cyano, amino, protected amino, nitro, optionally substituted —C₁-C₈alkyl, optionally substituted —C₁-C₈ alkoxy, optionally substituted—NHC₁-C₈ alkyl, optionally substituted —S—(—C₁-C₈ alkyl), optionallysubstituted —SO₂—(—C₁-C₈ alkyl), -optionally substituted —SO₂—NH—(—C₁-C₈alkyl), optionally substituted —NH—SO₂—(—C₁-C₈ alkyl), —CO₂R₁₂,—NR₁₃R₁₄, and —CO—NR₁₃R₁₄; R₁₂ is selected from the group consistingof: 1) Optionally substituted —C₁-C₈ alkyl; 2) Optionally substituted—C₂-C₈ alkenyl; 3) Optionally substituted —C₂-C₈ alkynyl; 4) Optionallysubstituted —C₃-C₈ cycloalkyl; 5) Optionally substituted —C₃-C₈cycloalkenyl; 6) Optionally substituted 3- to 8-memberedheterocycloalkyl; 7) Optionally substituted aryl; and 8) Optionallysubstituted heteroaryl; R₁₃ and R₁₄ are each independently selected fromhydrogen, optionally substituted —C₁-C₈-alkyl, optionally substituted—C₂-C₈-alkenyl, optionally substituted —C₂-C₈-alkynyl; optionallysubstituted —C₃-C₈-cycloalkyl; optionally substituted —C₃-C₈cycloalkenyl; optionally substituted 3- to 12-membered heterocycloalkyl,optionally substituted aryl; optionally substituted heteroaryl;optionally substituted —C₁-C₈-alkoxy, —C(O)R₁₂, —S(O)₂R₁₂, and—S(O)₂NHR₁₂; alternatively, R₁₃ and R₁₄ are taken together with thenitrogen atom to which they are attached to form an optionallysubstituted heterocyclic ring; R₄ is selected from hydrogen and —C₁-C₈alkyl; and n is 0 to k, wherein k is the total number of CH and NHgroups in A when A is unsubstituted.
 2. The compound of claim 1,represented by Formula (Ib) or a pharmaceutically acceptable saltthereof:

wherein R₁, R₂, R₃, R₄, A, B, L, X, and n are as defined in claim
 1. 3.The compound of claim 1, represented by Formula (III-1) or apharmaceutically acceptable salt thereof:

wherein R₁, R₂, R₃, R₄, A, B, L, and n are as defined in claim
 1. 4. Thecompound of claim 1, wherein A is selected from one of the following byremoval of hydrogen atoms from two adjacent ring carbon atoms:

wherein each of the above shown is optionally substituted with one ormore substituents which are not R₃ when possible.
 5. The compound ofclaim 1, wherein B is selected from one of the following by removal ofone or two hydrogen atoms:

wherein each of the above shown is optionally substituted when it ispossible.
 6. The compound of claim 1, wherein R₂ is selected from one ofthe following by removal of one ring hydrogen atom:

wherein each of the above shown is optionally substituted.
 7. Thecompound of claim 1, wherein R₂ is selected from the following:

wherein each group is optionally substituted.
 8. The compound of claim1, wherein R₂ is selected from the groups set forth in Table 1: TABLE 1Entry R₂ 1

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9. A compound selected from the compounds set forth below or apharmaceutically acceptable salt thereof: Com- pound Structure 1

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10. A pharmaceutical composition comprising a compound according toclaim 1, in combination with a pharmaceutically acceptable carrier,diluent or excipient.
 11. A method of treating or preventing an RSVinfection in a subject in need thereof, comprising administering to thesubject a therapeutically effective amount of a compound represented byFormula (I), or a pharmaceutically acceptable salt thereof:

wherein: A is selected from the group consisting of: 1) Optionallysubstituted heteroaryl; and 2) Optionally substituted aryl; B isselected from the group consisting of: 1) Optionally substitutedheteroaryl; and 2) Optionally substituted aryl; L is —NH—, —NHC(O)—, or—NHC(O)NH—; X is —CH₂—, or —C(O)—; R₁ is selected from the groupconsisting of: 1) Optionally substituted heteroaryl; and 2) Optionallysubstituted aryl; R₂ is absent or is selected from the group consistingof: 1) Optionally substituted —C₁-C₈ alkyl; 2) Optionally substituted—C₂-C₈ alkenyl; 3) Optionally substituted —C₂-C₈ alkynyl; 4) Optionallysubstituted —C₁-C₈ alkoxy; 5) Optionally substituted aryloxy; 6)Optionally substituted —C₃-C₁₂ cycloalkyl; 7) Optionally substituted—C₃-C₁₂ cycloalkenyl; 8) Optionally substituted 3- to 12-memberedheterocycloalkyl; 9) Optionally substituted aryl; 10) Optionallysubstituted arylalkyl; 11) Optionally substituted heteroaryl; 12)Optionally substituted heteroarylalkyl; 13) —NR₁₃R₁₄; 14) —CO—NR₁₃R₁₄;and 15) —SO₂—NR₁₃R₁₄; Each R₃ is independently selected from halogen,hydroxyl, protected hydroxyl, cyano, amino, protected amino, nitro,optionally substituted —C₁-C₈ alkyl, optionally substituted —C₁-C₈alkoxy, optionally substituted —NHC₁-C₈ alkyl, optionally substituted—S—(—C₁-C₈ alkyl), optionally substituted —SO₂—(—C₁-C₈ alkyl),-optionally substituted —SO₂—NH—(—C₁-C₈ alkyl), optionally substituted—NH—SO₂—(—C₁-C₈ alkyl), —CO₂R₁₂, —NR₁₃R₁₄, and —CO—NR₁₃R₁₄; R₁₂ isselected from the group consisting of: 1) Optionally substituted —C₁-C₈alkyl; 2) Optionally substituted —C₂-C₈ alkenyl; 3) Optionallysubstituted —C₂-C₈ alkynyl; 4) Optionally substituted —C₃-C₈ cycloalkyl;5) Optionally substituted —C₃-C₈ cycloalkenyl; 6) Optionally substituted3- to 8-membered heterocycloalkyl; 7) Optionally substituted aryl; and8) Optionally substituted heteroaryl; R₁₃ and R₁₄ are each independentlyselected from hydrogen, optionally substituted —C₁-C₈-alkyl, optionallysubstituted —C₂-C₈-alkenyl, optionally substituted —C₂-C₈-alkynyl;optionally substituted —C₃-C₈-cycloalkyl; optionally substituted —C₃-C₈cycloalkenyl; optionally substituted 3- to 12-membered heterocycloalkyl,optionally substituted aryl; optionally substituted heteroaryl;optionally substituted —C₁-C₈-alkoxy, —C(O)R₁₂, —S(O)₂R₁₂, and—S(O)₂NHR₁₂; alternatively, R₁₃ and R₁₄ are taken together with thenitrogen atom to which they are attached to form an optionallysubstituted heterocyclic ring; R₄ is selected from hydrogen andoptionally substituted —C₁-C₈ alkyl; and n is 0 to k, wherein k is thetotal number of CH and NH groups in A when A is unsubstituted.
 12. Themethod of claim 11, further comprising administering to the subject asteroid anti-inflammatory compound.
 13. The method of claim 11, furthercomprising the step of administering to the subject an additionalanti-RSV agent.
 14. The method of claim 11, further comprisingadministering to the subject an anti-influenza compound.
 15. The methodof claim 13, wherein the compound and the additional anti-RSV agent areco-formulated.
 16. The method of claim 13, wherein the compound and theadditional anti-RSV agent are co-administered.
 17. The compound of claim1, represented by Formula (III-2) or a pharmaceutically acceptable saltthereof:

wherein R₁, R₂, R₃, R₄, A, B, L, and n are as defined in claim
 1. 18.The compound of claim 1, represented by Formula (Ia) or apharmaceutically acceptable salt thereof:

wherein R₁, R₂, R₃, R₄, A, B, L, X, and n are as defined in claim 1.